Gene Synthesis & Cloning/Mutagenesis FAQs



The Most Commonly Asked Questions

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Service Service Offering  Estimated Turnaround Time  Price  
FragmentGENE Synthesis of double-stranded DNA fragments 2-4 business days  $
PriorityGENE Standard gene synthesis with cloning into a plasmid 8-10 business days  $$
TurboGENE 5 Expedited gene synthesis with cloning into a plasmid and our fastest available turnaround 5 business days  $$$
TurboGENE 7 Expedited gene synthesis with cloning into a plasmid 7 business days  $$$
Antibody DNA Synthesis  Synthesis of heavy and light chain sequences with cloning into a plasmid 6-8 business days  $$
AAV Plasmid Synthesis Synthesis of AAV sequences and cloning into a plasmid with AAV-ITR sequence verification using proprietary technologies  11-15 business days  $$$
CRISPR Construct Synthesis  Synthesis of gRNA and cloning into a custom plasmid 8-10 business days  $
ssDNA Synthesis Synthesis of single-stranded DNA fragments 10-15 business days  $$
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To access pricing information for your institution, please contact your sales representative.
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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.


Log in to your online GENEWIZ 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.

 

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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 here.

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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 GENEWIZ 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. 

GENEWIZ 
Attn: Project Management
115 Corporate Boulevard
South Plainfield, NJ 07080
USA 

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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 here.

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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. 

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GENEWIZ 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:

a. Spin the tube briefly to ensure that the contents are on the bottom of the tube

b. Resuspend the sample in a volume appropriate for your needs:

i. For example, 20-50 µL for ~0.25 to ~0.1 µg/µL of sterile high purity water

c. The resuspension solution is dependent on downstream applications - TE may interfere with various enzyme manipulations

d. Vortex briefly, let the DNA tube sit for 2-10 minutes on ice, then vortex again

 

 

 

ORDERING

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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. 

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. 
 

Upload Download SS


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To confirm a quote, log in to your online GENEWIZ account. All submitted inquiries and orders will be visible at the bottom of the page under “My Orders”. 
 
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.
 

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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 (PM@genewiz.com), or live chat. 

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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. 

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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.

SERVICE DETAILS

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Backed by an over 99.9% project completion rate, GENEWIZ 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. 
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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 pUC-GW vector; the insert is verified by Sanger DNA sequencing and restriction digestion.
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GENEWIZ will synthesize cloned gene sequences ranging from 1 bp to 50+ kb, using short DNA oligos as building blocks.

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GENEWIZ 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, GENEWIZ optimizes your gene for synthesis. 

In some very rare instances, extreme sequence content may be refractory to available sequence verification techniques. GENEWIZ 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.

 
A:

GENEWIZ 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.

 
A:

At GENEWIZ, the protection of our customers' intellectual property is a top priority. Proprietary information is always under client ownership, and GENEWIZ places the utmost importance on confidentiality and privacy. Comprehensive protective measures enable GENEWIZ to earn and maintain the trust and confidence of all customers, collaborators, and partners. For more information about the GENEWIZ Confidentiality Policy, please click here.

CODON OPTIMIZATION

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GENEWIZ 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.
 
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GENEWIZ’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:

a. Codon usage bias

b. GC content

c. mRNA secondary structure

d. Custom desired patterns

e. Custom undesired patterns

f. Repeat sequences (direct repeat, inverted repeat, and dyad repeat)

g. Restriction enzyme recognition sites (deletion or insertion)

Please note, GENEWIZ cannot guarantee final protein expression. 

 

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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 PM@genewiz.com.

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Yes, GENEWIZ’s codon optimization tool is available in the “My Tools” section within your online account and can be utilized at any time.

My Tools SS


VARIANT SYNTHESIS

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The Project Management Team typically creates the primary sequence and works on the variant constructs after the creation of the primary construct.
 
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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.

 

CLONING AND MUTAGENESIS

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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.


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GENEWIZ 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.
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GENEWIZ currently only offers cloning vectors based on the pUC-GW-Kan/Amp backbone.  We are unable to purchase proprietary vectors on your behalf.
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 Yes, the Project Management Team starts gene synthesis at the time of order confirmation.
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 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.
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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. 
A:

For any information regarding on-going promotional discounts, please contact your sales representative.

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  Site-Directed Mutagenesis Insert PCR-Based Mutagenesis  TurboMUTANT 
Plasmid Size Guidelines  10kb or less in size, 
Non-complex
> 10 kb/unstable plasmid  < 8 kb/stable plasmid
Turnaround Time Starts from 8-10 BD Starts from 10-15 BD  Starts from 5 BD 

QUALITY AND FINAL DELIVERABLES

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GENEWIZ 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, GENEWIZ 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.
 

 
A:

Standard deliverables for GENEWIZ gene synthesis projects include:

a. 2-5 µg of lyophilized plasmid containing your desired synthetic construct

b. Certificate of Analysis (COA), including restriction digest

c. Sequence trace data with alignment

d. Sequence of synthetic gene alone and in the designated vector


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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.


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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.


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The GENEWIZ 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.

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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). 

A:

Some extremely complex sequences are expected to not be verifiable using current Sanger sequencing techniques and are noted in the quote. In the very 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).  


How to Order Gene Synthesis

Have a specific question?

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