GENEWIZ | Gene Synthesis FAQs

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document, '//static.hotjar.com/c/hotjar-', '.js?sv='); </script>  <script src="https://cdn.cookielaw.org/scripttemplates/otSDKStub.js" type="text/javascript" charset="UTF-8" data-domain-script="0190a77c-46be-74e5-a65b-09fc07bbf439" ></script> <script type="text/javascript"> function OptanonWrapper() { } </script>   <p style="display:none;"><button id="ot-sdk-btn" class="ot-sdk-show-settings">CookieSettings</button></p>  <br /><br /> <div class="headline-center Rich-Text-Edit-Area"> <br /> <br /> <h2>Gene Synthesis & Cloning/Mutagenesis FAQs</h2> </div> <div class="headline-center Rich-Text-Edit-Area"> <div class="container"> <ul class="list-inline"> <li> <h3 style="text-align:left;"><a name="Commonly-Asked-Questions" href="#Commonly-Asked-Questions" class="blue-text">The Most Commonly Asked Questions</a></h3> </li> | <li> <h3 style="text-align:left;"><a name="Ordering" href="#Ordering" class="blue-text">Ordering</a></h3> </li> | <li> <h3 style="text-align:left;"><a name="Service-Details" href="#Service-Details" class="blue-text">Service Details</a></h3> </li> | <li> <h3 style="text-align:left;"><a name="Codon-Optimization" href="#Antibody-Production" class="blue-text">Recombinant Antibody Production</a></h3> </li> | <li> <h3 style="text-align:left;"><a name="Codon-Optimization" href="#Lentivirus" class="blue-text">Lentivirus</a></h3> </li> | <li> <h3 style="text-align:left;"><a name="Codon-Optimization" href="#Codon-Optimization" class="blue-text">Codon Optimization</a></h3> </li> | <li> <h3 style="text-align:left;"><a name="Cloning-and-Mutagenesis" href="#Cloning-and-Mutagenesis" class="blue-text">Cloning and Mutagenesis</a></h3> </li> | <li> <h3 style="text-align:left;"><a name="Quality-and-Final-Deliverables" href="#Quality-and-Final-Deliverables" class="blue-text">Quality and Final Deliverables</a></h3> </li> </ul> </div> <br /> <br /> </div> <div class="container FAQs"> <div class="row"> <span id="Commonly-Asked-Questions" class="anchor"></span> <p style="padding: 20px;"><b>The Most Commonly Asked Questions</b></p> </div> <div class="row"> <div class="NoBorder panel-group acc-v1 container" id='MOST-COMMONLY-ASKED-QUESTIONS'> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#MOST-COMMONLY-ASKED-QUESTIONS" href="#QA1" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What are the differences between your gene synthesis service options? </p></div> </a> </div> <div id="QA1" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <div class="table-column"> <table class="table table-responsive table-bordered"> <tbody class="panel panel-blue"> <tr class="panel-heading"> <td>Service</td> <td>Service Offering </td> <td>Estimated Turnaround Time </td> <td>Price  </td> </tr> <tr> <td>FragmentGENE</td> <td>Synthesis of double-stranded DNA fragments</td> <td>2-4 business days</td> <td> $</td> </tr> <tr> <td>PriorityGENE</td> <td style="vertical-align: middle;">Standard gene synthesis with cloning into a plasmid</td> <td>8-10 business days</td> <td> $$</td> </tr> <tr> <td>TurboGENE 5</td> <td style="vertical-align: middle;">Expedited gene synthesis with cloning into a plasmid and our fastest available turnaround</td> <td>5 business days</td> <td> $$$</td> </tr> <tr> <td>TurboGENE 7</td> <td style="vertical-align: middle;">Expedited gene synthesis with cloning into a plasmid</td> <td>7 business days</td> <td> $$$</td> </tr> <tr> <td>Antibody DNA Synthesis </td> <td style="vertical-align: middle;">Synthesis of heavy and light chain sequences with cloning into a plasmid</td> <td>6-8 business days</td> <td> $$</td> </tr> <tr> <td>AAV Plasmid Synthesis</td> <td style="vertical-align: middle;">Synthesis of AAV sequences and cloning into a plasmid with AAV-ITR sequence verification using proprietary technologies </td> <td>11-15 business days</td> <td> $$$</td> </tr> <tr> <td>CRISPR Construct Synthesis </td> <td style="vertical-align: middle;">Synthesis of gRNA and cloning into a custom plasmid</td> <td>8-10 business days</td> <td> $</td> </tr> <tr> <td>ssDNA Synthesis</td> <td style="vertical-align: middle;">Synthesis of single-stranded DNA fragments</td> <td>10-15 business days</td> <td> $$</td> </tr> </tbody> </table> </div> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#MOST-COMMONLY-ASKED-QUESTIONS" href="#QA2" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Where can I get general pricing before submitting a quote request? </p></div> </a> </div> <div id="QA2" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> To access pricing information for your institution, please contact your sales representative.<br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#MOST-COMMONLY-ASKED-QUESTIONS" href="#QA3" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>How do I request a quote for a gene synthesis service? </p></div> </a> </div> <div id="QA3" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>PriorityGENE, TurboGENE, ValueGENE, antibody DNA synthesis, and AAV plasmid synthesis services require you to request a quote before ordering. FragmentGENE, CRISPR construct synthesis, and ssDNA synthesis services are available for direct order, so you do not need to request a quote for these services.</p> <br /> Log in to your online Azenta account and select “Gene Synthesis”, then click on the name of the specific service you are interested in. You will be directed to a quote request form for that selected service. Once you complete the form, select the “Review Inquiry” button at the bottom of the screen. After you review the entered details, select the “Submit Quote Request” button to request your quote. Upon submission of the inquiry, you can expect to receive a quotation within one business day. You can view a PDF copy of the order information including price and download using the “Print Price” button. <p> </p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#MOST-COMMONLY-ASKED-QUESTIONS" href="#QA4" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What vector do you clone into?</p></div> </a> </div> <div id="QA4" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>The assembled full-length gene is cloned into the pUC-GW-Kan/Amp vector via the EcoRV site. The final construct is verified with both Sanger DNA sequencing (on at least one strand) and restriction digestion. The sequences of pUC-GW-Kan/Amp can be found <a href="https://3478602.fs1.hubspotusercontent-na1.net/hubfs/3478602/2022-02%20GENE%20SYNTH%20-%20Gene%20to%20Antibody/12034-M&G%200522%20pUC-GW%20Plasmid%20Map%20Guide.pdf">here</a>.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#MOST-COMMONLY-ASKED-QUESTIONS" href="#QA5" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>How do I submit the starting material required for cloning? </p></div> </a> </div> <div id="QA5" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>We currently process in-house pUC-GW-Kan/Amp plasmids at our facility free of charge. As such, we request that a 5 µg aliquot of circularized plasmid (min. concentration of 20 ng/µl) is provided to our facility. This sample can be directly shipped at room temperature to the address below, or it can be submitted via any available Azenta dropbox. To ensure the sample is correctly identified, please include the first page of the order receipt or quotation in your package. If those documents are not available, the project tracking number is acceptable as well. <br /> <br /> Azenta <br /> Attn: Project Management<br /> 115 Corporate Boulevard<br /> South Plainfield, NJ 07080<br /> USA </p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#MOST-COMMONLY-ASKED-QUESTIONS" href="#QA6" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Can you store my vector?</p></div> </a> </div> <div id="QA6" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p> We store any starting material provided and final constructs generated at our facility for up to two years to be used for future orders. For more information, please find our Sample Storage Policy <a href="https://www.genewiz.com/en/Public/Resources/Sample-Submission-Guidelines/Sample-Storage-Policy-Sample-Submission-Guidelines">here</a>. </p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#MOST-COMMONLY-ASKED-QUESTIONS" href="#QA7" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What timeline should I expect for my project? </p></div> </a> </div> <div id="QA7" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p> The timeline will depend on the size of the gene to be synthesized, the complexity of the vector for cloning, and the requested DNA preparation scale. For reference, the turnaround time for a gene less than 1.5 kb is 8-10 business days with our PriorityGENE service, 7 business days with TurboGENE 7 priority, or 5 business days with TurboGENE 5 priority. A more specific turnaround time for each order is included within the quotation, as well as the “Order Review” page within your online account. </p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#MOST-COMMONLY-ASKED-QUESTIONS" href="#QA8" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is the recommended DNA resuspension protocol? </p></div> </a> </div> <div id="QA8" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Azenta sample tubes are delivered containing 2-5 µg lyophilized plasmid DNA for gene synthesis orders, or 500 ng - 1 µg purified linear dsDNA for FragmentGENE orders. For resuspension, we recommend the following:</p> <p style="text-indent: 40px;">a. Spin the tube briefly to ensure that the contents are on the bottom of the tube</p> <p style="text-indent: 40px;">b. Resuspend the sample in a volume appropriate for your needs:</p> <p style="text-indent: 70px;">i. For example, 20-50 µL for ~0.25 to ~0.1 µg/µL of sterile high purity water</p> <p style="text-indent: 40px;">c. The resuspension solution is dependent on downstream applications - TE may interfere with various enzyme manipulations</p> <p style="text-indent: 40px;">d. Vortex briefly, let the DNA tube sit for 2-10 minutes on ice, then vortex again</p> <p> </p> <p> </p> <div> </div> </div> </div> </div> </div> </div> </div><div class="container FAQs"> <div class="row"> <span id="Ordering" class="anchor"></span> <p style="padding: 20px;"><b>ORDERING</b></p> </div> <div class="row"> <div class="NoBorder panel-group acc-v1 container" id='ORDERING-FAQ'> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#ORDERING-FAQ" href="#QA-1" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>How do I request a bulk gene synthesis order?</p></div> </a> </div> <div id="QA-1" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <div class="table-column"> If you have multiple sequences within your order, we recommend using the “Grid View” option available at the top right corner of the online order forms. Using this format, you can select the arrow at the top of each column to autofill any identical information within a column, similar to an Excel sheet. <br /> <br /> Alternatively, please feel free to download our Ordering Template and fill out the necessary information within this form. To access this form within a gene synthesis inquiry, select the “Download/Upload” button located at the top right corner of the screen. After this is done, select the same “Download/Upload” button to view the option to upload the completed form. <br /> <div> </div> </div> <style type="text/css"> .protocol-3 { width: 1170px; display: inline-block; text-align: left; } .protocol-3 img { width: 30%; height: auto; display: block; margin-left: auto; margin-right: auto; } @media screen and (max-width: 1170px) { .protocol-1 { width: auto; } } </style> <div class="protocol-3"> <div class="protocol-3 img"> <h3 style="text-align: center;"> </h3> <a rel="noopener noreferrer" rel="noopener noreferrer" target="_blank"><img alt="Upload Download SS" src="/-/media/Images/Upload-Download-SS.ashx?la=en&hash=AA9110F757DD9616930E57011B237D7C5F00B324" /></a></div> <br /> <br /> </div> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#ORDERING-FAQ" href="#QA-2" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>How do I confirm a gene synthesis quote (PriorityGENE, TurboGENE, ValueGENE, antibody DNA synthesis)? </p></div> </a> </div> <div id="QA-2" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> To confirm a quote, log in to your online Azenta account. All submitted inquiries and orders will be visible at the bottom of the page under “My Orders”. <br />  <br /> Select the “Review Quote” button next to the related project tracking number (30-xxxxxxxxx). After this is done, an “Add to Cart” button will appear at the bottom of the screen. Once this is selected, you will be redirected to the ordering page where you will be able to review our Terms and Conditions and enter the payment information for this order. After this is done, select the “Confirm” button at the bottom of the screen to place your order.<br /> <div> </div> <br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#ORDERING-FAQ" href="#QA-3" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Who provides support for gene synthesis projects?</p></div> </a> </div> <div id="QA-3" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p> Our Project Management Team is happy to help with any questions. Please feel free to contact us from 9:00 AM - 6:30 PM EST via phone (+1-908-222-0711 ext. 3), email (<a href="mailto:PM@azenta.com">PM@azenta.com</a>), or live chat. </p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#ORDERING-FAQ" href="#QA-4" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is the advantage of using the antibody DNA synthesis service vs. PriorityGENE? </p></div> </a> </div> <div id="QA-4" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p> Our antibody DNA synthesis service provides synthesis and cloning of your antibody heavy/light chain sequences up to 1.5 kb into any custom vector in as few as 6 business days, the fastest turnaround time on the market. Our PriorityGENE service provides synthesis, assembly, and cloning of sequences with unlimited length on a timely basis. </p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#ORDERING-FAQ" href="#QA-5" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is the advantage of using the AAV plasmid synthesis service? </p></div> </a> </div> <div id="QA-5" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p> With our AAV plasmid synthesis service, we can synthesize and clone transgene expression cassettes into custom AAV vectors with high efficiencies. All final products will come bundled with mini-scale or large-scale DNA preparation using our new AAV plasmid preparation protocol and AAV-ITR sequence verified AAV plasmids. This service also offers ITR correction if mutations are found after sequence-verification of these regions.</p> </div> </div> </div> </div> </div> </div><div class="container FAQs"> <div class="row"> <span id="Service-Details" class="anchor"></span> <p style="padding: 20px;"><b>SERVICE DETAILS</b></p> </div> <div class="row"> <div class="NoBorder panel-group acc-v1 container" id='SERVICE-DETAILS'> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#SERVICE-DETAILS" href="#QA---1" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What are the benefits of choosing your gene synthesis service? </p></div> </a> </div> <div id="QA---1" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <div class="table-column"> Backed by an over 99.9% project completion rate, Azenta can synthesize naturally occurring sequences, codon-optimized genes, gene libraries, complex sequences, large sequences, or any other sequence you need for your research - regardless of complexity. </div> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#SERVICE-DETAILS" href="#QA---2" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>How do you synthesize genes?</p></div> </a> </div> <div id="QA---2" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> Single-stranded oligonucleotides are designed, synthesized, and assembled using different protocols depending on sequence characteristics. Subsequently, the assembled full-length gene is cloned into the <a href="https://cdn2.hubspot.net/hubfs/3478602/GENEWIZ_pUC-GW_Plasmid_Map.pdf">pUC-GW </a>vector; the insert is verified by Sanger DNA sequencing and restriction digestion.<br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#SERVICE-DETAILS" href="#QA---3" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What gene length can you synthesize?</p></div> </a> </div> <div id="QA---3" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p> Azenta will synthesize cloned gene sequences ranging from 1 bp to 50+ kb, using short DNA oligos as building blocks. </p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#SERVICE-DETAILS" href="#QA---4" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Can you synthesize sequences with high GC content or repeats?</p></div> </a> </div> <div id="QA---4" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Azenta has a proven track record of synthesizing difficult templates, such as GC-rich sequences and sequences with repeats, using our proprietary technologies. With our bioinformatics platform, Azenta optimizes your gene for synthesis. <br /> <br /> In some very rare instances, extreme sequence content may be refractory to available sequence verification techniques. Azenta carefully reviews every sequence to estimate difficulty and anticipate project challenges and customizes every project quotation to include an estimated turnaround time and pricing information based on this analysis.</p> <div> </div> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#SERVICE-DETAILS" href="#QA---5" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Are there any additional fees beyond the per bp price? </p></div> </a> </div> <div id="QA---5" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Azenta provides a custom quote for every gene synthesis project. For standard priority, typical gene sequences (non-repetitive, average GC-content) that do not require custom subcloning, no additional fees will apply. For genes with expedited handling, difficult sequences, large inserts, or for projects with custom subcloning requirements, any relevant fees will be indicated in your custom gene synthesis quote.</p> <div> </div> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#SERVICE-DETAILS" href="#QA---6" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is your policy for intellectual property protection? </p></div> </a> </div> <div id="QA---6" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p> At Azenta, the protection of our customers' intellectual property is a top priority. Proprietary information is always under client ownership, and Azenta places the utmost importance on confidentiality and privacy. Comprehensive protective measures enable Azenta to earn and maintain the trust and confidence of all customers, collaborators, and partners. For more information about the Azenta Confidentiality Policy, please <a href="https://www.genewiz.com/en/Public/Company/Policies/Confidentiality-Policy">click here</a>.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#SERVICE-DETAILS" href="#QA---7" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>How does variant synthesis work? Do you work on the synthesis in parallel or sequential order? </p></div> </a> </div> <div id="QA---7" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <div class="table-column"> The Project Management Team typically creates the primary sequence and works on the variant constructs after the creation of the primary construct.<br /> <div> </div> </div> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#SERVICE-DETAILS" href="#QA---8" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Why is the cost of variant synthesis different from the other genes within my order? </p></div> </a> </div> <div id="QA---8" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>As the variants are being created from the parent sequence, we can offer a discounted price for variant synthesis. Please keep in mind, variant synthesis does add an additional 3-5 business days to the quoted turnaround time.</p> <div> </div> <br /> </div> </div> </div> </div> </div> </div><div class="container FAQs"> <div class="row"> <span id="Antibody-Production" class="anchor"></span> <p style="padding: 20px;"><strong>RECOMBINANT ANTIBODY PRODUCTION</strong></p> </div> <div class="row"> <div class="NoBorder panel-group acc-v1 container" id='RECOMBINANT-ANTIBODY-PRODUCTION'> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#RECOMBINANT-ANTIBODY-PRODUCTION" href="#1---QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Do you offer any expression vectors?</p></div> </a> </div> <div id="1---QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Yes, we offer a pETE vector which is optimized for our recombinant expression platform. You can provide your own preferred expression vector as well.</p> <div></div> <br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#RECOMBINANT-ANTIBODY-PRODUCTION" href="#2---QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Where can I get the antibody genes? </p></div> </a> </div> <div id="2---QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> Azenta can synthesize and clone your genes of interest or you can provide your own DNA constructs.<br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#RECOMBINANT-ANTIBODY-PRODUCTION" href="#3---QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What cell lines are available for expression?</p></div> </a> </div> <div id="3---QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> We offer a number of mammalian cell lines including CHO-S, HEK-293, ExpiCHO/Expi293 and a variety of bacterial (<em>E.coli</em>) strains for antibody expression.<br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#RECOMBINANT-ANTIBODY-PRODUCTION" href="#4---QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Can you express more than antibodies? </p></div> </a> </div> <div id="4---QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> Yes, we can express your antibodies (IgGs), bispecific formats, and antibody fragments (ScFv, VHH, Fab).<br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#RECOMBINANT-ANTIBODY-PRODUCTION" href="#5---QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is the maximum project throughput and scale?</p></div> </a> </div> <div id="5---QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> Antibodies are expressed in high-throughput (up to 500 unique antibodies) at mini/midi scales up to 1L to deliver ug to mg quantities of purified material.<br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#RECOMBINANT-ANTIBODY-PRODUCTION" href="#6---QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is the quality control process for purified antibodies?</p></div> </a> </div> <div id="6---QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> All projects are analyzed by A280 for antibody concentration and SDS-PAGE for purity assessment. Common optional QC add-ons include aSEC, endotoxin analysis, and mass spectrometry services, and high-throughput ELISA. Additional analyses are available, inquire for any custom QC requests.<br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#RECOMBINANT-ANTIBODY-PRODUCTION" href="#7---QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Are affinity characterization assessments available?</p></div> </a> </div> <div id="7---QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Yes, we perform antibody affinity characterization by surface plasmon resonance (SPR) and bio-layer interferometry (BLI).</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#RECOMBINANT-ANTIBODY-PRODUCTION" href="#8---QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What if some of my plasmids do not express an antibody? </p></div> </a> </div> <div id="8---QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Expression analysis is performed by BLI (Octet) to estimate the amount of protein in the supernatant. Non-producing plasmids will be reported to the customer and excluded from purification, so you do not pay purification fees on non-producing constructs. </p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#RECOMBINANT-ANTIBODY-PRODUCTION" href="#9---QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What final product can I expect to receive from you? </p></div> </a> </div> <div id="9---QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Standard deliverables include: 2-5ug synthesized DNA construct (if performed by Azenta), antibodies provided as purified protein or supernatant, Certificate of Analysis (CoA)</p> </div> </div> </div> </div> </div> </div><div class="headline-center Rich-Text-Edit-Area"> </div> <div class="container FAQs"> <div class="row"> <span id="Lentivirus" class="anchor"></span> <p style="padding: 20px;"><strong>LENTIVIRUS</strong></p> </div> <div class="row"> <div class="NoBorder panel-group acc-v1 container" id='LENTIVIRUS'> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#LENTIVIRUS" href="#QA----1" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is lentivirus? </p></div> </a> </div> <div id="QA----1" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Lentivirus, meaning “slow virus”, is a genus of retrovirus. Retroviruses express reverse transcriptase to convert their RNA genome into double-stranded DNA (dsDNA) and integrase to incorporate this viral DNA into the host’s genome. Due to this permanent DNA integration, the host cell produces more retroviruses as it divides, resulting in long-term expression of the disease.</p> <p>Lentivirus has long incubation periods between host infection and the onset of disease symptoms, such as human immunodeficiency virus (HIV) in the case of human acquired immune deficiency syndrome (AIDS). In biotechnological applications, recombinant lentiviral vectors are modified for safe handling and used as a gene delivery vector for mammalian cells.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#LENTIVIRUS" href="#QA----2" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is the advantage of using lentivirus? </p></div> </a> </div> <div id="QA----2" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Unlike other retroviruses that only infect dividing cells, lentiviruses infect both dividing and non-dividing (postmitotic) cells. This broad tropism, coupled with the stable integration into a host cell genome, allows for diverse <em>in vitro</em> and <em>in vivo</em> applications, including clinical cell therapy and CAR-T cell engineering. Additional advantages of lentivirus include the ability to express large transgenes (up to 9 kb) and low immunogenicity.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#LENTIVIRUS" href="#QA----3" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>How do your produce lentivirus? </p></div> </a> </div> <div id="QA----3" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Our team of experts uses HEK293T cells for lentiviral vector production. Following successful transfection with four packaging plasmids, the cell machinery facilitates the assembly and replication of the lentiviral particles during an approximately three-day incubation. Lentivirus particles are then harvested, filtered, purified, concentrated, and aliquoted to small volumes to facilitate your downstream transduction applications.</p> <p><img alt="Lentivirus Diagram" src="/-/media/12003-SD-0224-Lentivirus-Diagram-V2.ashx?h=441&w=800&la=en&hash=8FF673951BF323AAC3EB3C12F3117C6D0D6A6DDD" style="height: 441px; width: 800px;" /></p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#LENTIVIRUS" href="#QA----4" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What safety concerns should I be aware of when working with lentivirus?</p></div> </a> </div> <div id="QA----4" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Recombinant lentiviral vectors are generally considered safe as they are engineered to be replication incompetent, with the third-generation considered the safest. It features a chimeric 5’ LTR, self-inactivating truncated 3’ LTR, elimination of Tat, and a four-plasmid transfection system that has a low likelihood of generating a replication-competent virus. According to the guidelines of the National Institutes of Health (NIH), lentiviral particles should be handled as Risk Group 2 (RG2). Our team uses the third-generation packaging system in biosafety level 2 (BSL2) facilities for all default lentiviral handling.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#LENTIVIRUS" href="#QA----5" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Can lentivirus vectors be packaged with customized envelope proteins? </p></div> </a> </div> <div id="QA----5" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Yes! Upon review, our expert team can accommodate custom requests. Our default option is an envelope glycoprotein of vesicular stomatitis virus (VSV-G protein). </p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#LENTIVIRUS" href="#QA----6" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Can pooled libraries be packaged into lentivirus particles?</p></div> </a> </div> <div id="QA----6" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Yes! Our team of experts have extensive experience packaging various library types, including combinatorial and trimer-controlled libraries. The diversity of the final packaged lentivirus will depend on the plasmid material. If you do not already have a library sample, we can generate and validate this for you. Please see <a href="https://www.genewiz.com/en-GB/Public/Services/Gene-Synthesis/Synthetic-DNA-Libraries">here</a> for more information.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#LENTIVIRUS" href="#QA----7" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Do you offer transfer plasmids for cloning? </p></div> </a> </div> <div id="QA----7" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p> Yes! We offer various transfer plasmid options for your cloning needs in the context of lentivirus production. If we don't have a vector that suits your requirements, we can synthesize any necessary components to create a transfer plasmid that aligns with your design.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#LENTIVIRUS" href="#QA----8" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>How is quality control (QC) performed on lentivirus particles? </p></div> </a> </div> <div id="QA----8" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>We offer two QC options for the validation of your lentiviral particles:</p> <p style="text-indent: 40px;">1. <strong>p24 ELISA (physical titer)</strong>: This method, offered as our standard QC, utilizes a sandwich ELISA to detect the presence of p24 protein, a major capsid protein encoded by the Gag gene. The p24 protein level is measured spectrophotometrically against a standard curve. Next, the quantified p24 value is correlated to the virus particle number. To facilitate downstream applications, we convert the lentiviral particles (LP) to TU/mL by dividing by 100 (there are ~100-1000 LPs for every active virus). This method is considered as a measure of physical titer.</p> <p style="text-indent: 40px;">2. <strong>WPRE qPCR (functional titer)</strong>: An aliquot of the packaged lentivirus vector is used to transduce HEK293T cells at various dilution levels. After a two-day incubation period, the genomic DNA of the transduced cells is harvested. Following that, qPCR amplification of the WPRE element is then performed alongside a calibrated standard curve. The WPRE element is absent in wild-type HEK293T cells but exists within the lentiviral cassette of a transfer plasmid. As the WPRE elements can only be detected in HEK293T cells if transduction is successful, this method is considered as a measure of functional titer.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#LENTIVIRUS" href="#QA----9" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is the difference between physical and functional titer? </p></div> </a> </div> <div id="QA----9" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Physical titering approximates the total amount of virus present in the sample by correlating it to the spectrophotometrically quantified p24 in an ELISA assay, regardless of whether it possesses the gene of interest or ability to infect cells. </p> <p>Functional titer is a measure of virus infectivity, quantifying the copy number of the WPRE element, which is detectable only upon a successful viral DNA integration into the host cell genome. Generally, functional titer is 100-1000-fold lower than physical titer. We report physical titer after accounting for this ratio, and express p24 in TU/mL.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#LENTIVIRUS" href="#QA----10" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is pseudotyping? </p></div> </a> </div> <div id="QA----10" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Pseudotyping is the process of incorporating envelope glycoproteins from other viruses to generate lentivirus particles with altered tropism. This is often used to enhance the infectivity and stability of lentivirus. For example, VSV-G is commonly used for pseudotyping due to its broad tropism.</p> </div> </div> </div> </div> </div> </div><div class="container FAQs"> <div class="row"> <span id="Codon-Optimization" class="anchor"></span> <p style="padding: 20px;"><strong>CODON OPTIMIZATION</strong></p> </div> <div class="row"> <div class="NoBorder panel-group acc-v1 container" id='CORDON-OPTIMIZATION'> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CORDON-OPTIMIZATION" href="#QA----1" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Can you optimize genes? </p></div> </a> </div> <div id="QA----1" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <div class="table-column"> Azenta can deliver codon-optimized genes, designed to meet your exact research specifications. These sequences are always available for your review prior to confirming your order and are typically included in the project quote.<br /> <div> </div> </div> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CORDON-OPTIMIZATION" href="#QA----2" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>How does codon optimization enhance gene synthesis projects? </p></div> </a> </div> <div id="QA----2" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Azenta’s codon optimization tool can optimize for multiple critical parameters to stabilize DNA fragments and improve gene expression efficiency. Parameters include, but are not limited to:</p> <p style="text-indent: 40px;">a. Codon usage bias</p> <p style="text-indent: 40px;">b. GC content</p> <p style="text-indent: 40px;">c. mRNA secondary structure</p> <p style="text-indent: 40px;">d. Custom desired patterns</p> <p style="text-indent: 40px;">e. Custom undesired patterns</p> <p style="text-indent: 40px;">f. Repeat sequences (direct repeat, inverted repeat, and dyad repeat)</p> <p style="text-indent: 40px;">g. Restriction enzyme recognition sites (deletion or insertion)</p> <p>Please note, Azenta cannot guarantee final protein expression. </p> <div> </div> <br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CORDON-OPTIMIZATION" href="#QA----3" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Can you optimize my sequence for more than one expression system? </p></div> </a> </div> <div id="QA----3" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Yes, we offer dual optimization for any two expression systems requested.  Additional requirements are also considered for review by the project team. Please note that any dual-system optimization represents a compromise, and a sequence specifically optimized for any given system may outperform the compromise sequence. To add this to your order, please add the request to the “Order Comments” section of your gene synthesis inquiry or email the Project Management Team at <a href="mailto:gs@Azenta.com">gs@azenta.com</a>.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CORDON-OPTIMIZATION" href="#QA----4" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Can I codon optimize a sequence without submitting a quote request? </p></div> </a> </div> <div id="QA----4" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Yes, Azenta’s codon optimization tool is available in the “My Tools” section within your online account and can be utilized at any time.</p> <style type="text/css"> .protocol-1 { width: 1170px; display: inline-block; text-align: left; } .protocol-1 img { width: 60%; height: auto; display: block; margin-left: auto; margin-right: auto; } @media screen and (max-width: 1170px) { .protocol-1 { width: auto; } } </style> <div class="protocol-1"> <div class="protocol-1 img"> <h3 style="text-align: center;"> </h3> <a rel="noopener noreferrer" rel="noopener noreferrer" target="_blank"><img alt="My Tools SS" src="/-/media/Images/My-Tools-SS.ashx?la=en&hash=DCDF38C0AF6F31B9578BD25F7B2A75C85A589482" /></a></div> <br /> <br /> </div> </div> </div> </div> </div> </div> </div><div class="container FAQs"> <div class="row"> <span id="Cloning-and-Mutagenesis" class="anchor"></span> <p style="padding: 20px;"><b>CLONING AND MUTAGENESIS</b></p> </div> <div class="row"> <div class="NoBorder panel-group acc-v1 container" id='CLONING-AND-MUTAGENESIS'> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CLONING-AND-MUTAGENESIS" href="#2-QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>My vector is a commonly used expression vector. Do you have it in stock?</p></div> </a> </div> <div id="2-QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>We currently only provide our in-house pUC-GW-Kan/Amp vectors for cloning. If you’d prefer cloning into a different vector, an aliquot of your custom vector will need to be provided upon confirmation of your order.</p> <div></div> <br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CLONING-AND-MUTAGENESIS" href="#3-QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>If I don’t have the required amount of material for my project, what are my alternatives? </p></div> </a> </div> <div id="3-QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> Azenta can provide transformation and mini-scale plasmid preparation services to create the required template amount for your project. This can be requested as a separate DNA preparation order or can be bundled with your gene synthesis project. If the latter option is suitable, please add an order comment when submitting your gene synthesis inquiry.<br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CLONING-AND-MUTAGENESIS" href="#4-QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>I don’t have a custom vector for my project; can you purchase it for me? </p></div> </a> </div> <div id="4-QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> Azenta currently only offers cloning vectors based on the pUC-GW-Kan/Amp backbone.  We are unable to purchase proprietary vectors on your behalf.<br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CLONING-AND-MUTAGENESIS" href="#5-QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>If I cannot provide the starting material at the time of order confirmation, will the gene synthesis steps still be initiated? </p></div> </a> </div> <div id="5-QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div>  Yes, the Project Management Team starts gene synthesis at the time of order confirmation.<br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CLONING-AND-MUTAGENESIS" href="#6-QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Can you clone into my vector using Gibson assembly®?</p></div> </a> </div> <div id="6-QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div>  We currently offer an in-house recombinational method of cloning that is similar to Gibson assembly. This method can be requested within the order comments of the inquiry form.<br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CLONING-AND-MUTAGENESIS" href="#7-QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is the difference between subcloning and PCR cloning? </p></div> </a> </div> <div id="7-QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> Both services offer cloning of an insert from the existing template plasmid into a different destination plasmid. For our subcloning service, the insert to be cloned is already flanked by the restriction sites to be used for cloning. This service is for simple restriction/ligation. The PCR cloning service is offered when there is an additional PCR step required prior to cloning. This could be necessary to add the correct restriction sites, for example. Both options can be requested via the same “Subcloning” form, the Project Management Team will internally determine the best strategy upon receipt of your inquiry. <br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CLONING-AND-MUTAGENESIS" href="#8-QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Are there any discounts available for these services?</p></div> </a> </div> <div id="8-QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>For any information regarding on-going promotional discounts, please contact your sales representative.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#CLONING-AND-MUTAGENESIS" href="#9-QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What is the difference between site-directed mutagenesis, insert PCR-based mutagenesis, and TurboMUTANT? </p></div> </a> </div> <div id="9-QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <div class="table-column"> <table class="table table-responsive table-bordered"> <tbody class="panel panel-blue"> <tr class="panel-heading"> <td> </td> <td>Site-Directed Mutagenesis</td> <td>Insert PCR-Based Mutagenesis </td> <td>TurboMUTANT </td> </tr> <tr> <td>Plasmid Size Guidelines </td> <td>10kb or less in size, <br /> Non-complex</td> <td>> 10 kb/unstable plasmid</td> <td> < 8 kb/stable plasmid</td> </tr> <tr> <td>Turnaround Time</td> <td style="vertical-align: middle;">Starts from 8-10 BD</td> <td>Starts from 10-15 BD </td> <td>Starts from 5 BD </td> </tr> </tbody> </table> </div> </div> </div> </div> </div> </div> </div><div class="container FAQs"> <div class="row"> <span id="Quality-and-Final-Deliverables" class="anchor"></span> <p style="padding: 20px;"><b>QUALITY AND FINAL DELIVERABLES</b></p> </div> <div class="row"> <div class="NoBorder panel-group acc-v1 container" id='QUALITY-AND-FINAL-DELIVERABLES'> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#QUALITY-AND-FINAL-DELIVERABLES" href="#1--QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>How do I know that I am getting the correct sequence? </p></div> </a> </div> <div id="1--QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <div class="table-column"> Azenta conducts sequence verification during the gene synthesis process to ensure your synthesis product is as intended. Final plasmid constructs are clonally isolated and confirmed by Sanger DNA sequence analysis to at least single-strand depth across the synthesized insert. Purified PCR fragments are sequence-verified on both strands, although verification at the fragment ends is limited to single-strand depth. Subassemblies and intermediate products may be cloned and verified to contain the correct sequence, as needed. DNA sequence re-verification after scale-up DNA preparation is available as an added option to your gene synthesis order. On completion of your synthetic gene, Azenta delivers an electronic data package in parallel with your construct. The data package includes the original sequencing data files from construct verification to facilitate your independent review.<br /> <div> </div> <br /> <div> </div> </div> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#QUALITY-AND-FINAL-DELIVERABLES" href="#2--QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>How do you deliver synthetic genes?</p></div> </a> </div> <div id="2--QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Standard deliverables for Azenta gene synthesis projects include:</p> <p style="text-indent: 40px;">a. 2-5 µg of lyophilized plasmid containing your desired synthetic construct</p> <p style="text-indent: 40px;">b. Certificate of Analysis (COA), including restriction digest</p> <p style="text-indent: 40px;">c. Sequence trace data with alignment</p> <p style="text-indent: 40px;">d. Sequence of synthetic gene alone and in the designated vector</p> <br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#QUALITY-AND-FINAL-DELIVERABLES" href="#3--QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What controls are used for each step completed in my project? </p></div> </a> </div> <div id="3--QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>All reagents, material used, and equipment are kept under strict quality controls by the lab. The Project Management Team may decide to implement additional controls if an uncommon failure rate is observed.</p> <br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#QUALITY-AND-FINAL-DELIVERABLES" href="#4--QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>My project is under mutation correction, what does that mean?</p></div> </a> </div> <div id="4--QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Mutations can be found at multiple stages of a project. For example, a synthesized primer may contain an incorrect base. Mutation correction may delay delivery by 1-4 business days depending on the situation.</p> <br /> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#QUALITY-AND-FINAL-DELIVERABLES" href="#5--QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Can I add a construct to my already confirmed project? </p></div> </a> </div> <div id="5--QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>The Azenta Project Management Team is happy to address your additional project requests through a new quotation request that references the current open project. Where possible, any discounts applied to the current project will be incorporated into the new quotation.  Please note that you must confirm the new project within the ongoing project timeline in order to take advantage of these discounts.</p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#QUALITY-AND-FINAL-DELIVERABLES" href="#6--QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>Do you sequence verify all gene synthesis products? </p></div> </a> </div> <div id="6--QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Yes, unless otherwise specified in the quotation due to extreme complexity. Single-strand sequence verification across the entire insert occurs for every synthesis product.  If the sequence-verified synthesis product is associated with a large-scale preparation, the product from the large-scale plasmid preparation is sequenced across junctions.  Additional sequence verification on the second strand, or after large-scale plasmid preparation, is available if requested (additional charges apply). </p> </div> </div> </div> <div class="NoBorder panel panel-default"> <div class="panel-heading SingleBorder" style="text-align:left;"> <a class="accordion-toggle" data-toggle="collapse" data-parent="#QUALITY-AND-FINAL-DELIVERABLES" href="#7--QA" style="color:gray;text-align:left;"> <div class="QA col-sm-1">Q:</div> <div><p>What do you guarantee in cases where the construct cannot be sequence-verified or delivered? If the project cannot be completed, what am I expected to pay? </p></div> </a> </div> <div id="7--QA" class=" panel-collapse collapse NoBorder"> <div class="panel-body NoBorder"> <div class="QA col-sm-1">A:</div> <p>Some extremely complex sequences are expected to not be verifiable using current Sanger sequencing techniques and are noted in the quote. In the <em>very</em> rare event where the Project Management Team is not able to deliver the intended material, there will be no charges for your project (unless otherwise specified in the quotation due to extreme complexity).  </p> <br /> </div> </div> </div> </div> </div> </div><script language="javascript" type="text/javascript"> jQuery(document).ready(function ($) { $(".SingleIcon").find("img").addClass("img-responsive center-block"); }); </script> <div class="process-v1 icon-nav Component-BackGround-Default margin-top-0 margin-bottom-30 padding-top-10 padding-bottom-10"> <div class="container"> <div class="headline-center"> <br> <br> <h1>Azenta Service FAQs</h1>  <p></p> </div> <ul class="list-unstyled row row-centered"> <li class='col-md-3 col-sm-6 col-centered'> <a href='/Public/Resources/FAQs/FAQs-Molecular-Genetics' target=''> <div class='SingleIcon headline-center Component-BackGround-Default margin-top-0 margin-bottom-30 padding-top-10 padding-bottom-10'> <div class="row"> 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GENEWIZ | Gene Synthesis FAQs