The Most Commonly Asked Questions
|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||$$$|
|CRISPR Construct Synthesis||Synthesis of gRNA and cloning into a custom plasmid||8-10 business days||$|
|ssDNA Synthesis||Synthesis of single-stranded DNA fragments||10-15 business days||$$|
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.
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.
Please submit all required starting material to Azenta. For UK projects (40-) samples should be submitted to our UK subsidiary, and for continental Europe projects (50-/90-) samples should be submitted to our German subsidiary (addresses below) as soon as possible after initiating your order (if required). A detailed list of the required starting material is included in section III "How to start your order" of your gene synthesis quotation.
Kindly submit a 5µg aliquot (minimum concentration of 20ng/µL) of the undigested, purified, circular plasmid. Please ensure that the name of the plasmid on the tube matches the name submitted for the order. Include the first page of the order receipt or quote with the submitted plasmid sample to ensure they are accurately identified
For UK: DNASeqUK@genewiz.com | (+44 (0) 1279 873837)
For EU: Logistics.Europe@genewiz.com | (+49-341 520 122 - 41)
All vectors submitted to our facility will be held in storage for two years and available within this time for future project use.
Direct Shipping (FedEx, UPS & DHL):
If a Azenta dropbox is not available, or if your starting material requires dry ice, please use an overnight delivery service such as FedEx, UPS or DHL.
UK Shipping Address:
Attn: Project Management
Hope End, Takeley
Essex CM22 6TA
Tel: +44 01279 873837
Germany Shipping Address:
Attn: Project Management
Phone: +49-341 520 122-41
Azenta dropbox (If Available):
Starting material that is stable at room temperature may be submitted via a GENEWIZ dropbox (when available). Contact firstname.lastname@example.org (UK) or Logistics.Europe@genewiz.com (continental Europe) to request a dropbox pick-up for your location, or if you have any related questions. Please click here for other dropbox options.
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.
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.
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:
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
How do I confirm a gene synthesis quote (PriorityGENE, TurboGENE, ValueGENE, antibody DNA synthesis)?
Our Project Management Team is happy to help with any questions. Please feel free to contact us from 9:00 AM - 6:00 PM CET via phone (+49 (0) 341 520 122-41), email (PM@azenta.com), or live chat.
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.
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.
Azenta can synthesize cloned gene sequences ranging from 1 bp to 50+ kb, using short DNA oligos as building blocks.
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.
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.
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.
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 click here.
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.
RECOMBINANT ANTIBODY PRODUCTION
Yes, we offer a pETE vector which is optimized for our recombinant expression platform. You can provide your own preferred expression vector as well.
Azenta can synthesize and clone your genes of interest, or you can provide your own DNA constructs.
We offer a number of mammalian cell lines including CHO-S, HEK-293, ExpiCHO/Expi293, and a variety of bacterial (E.coli) strains for antibody expression.
Yes, we can express your antibodies (IgGs), bispecific formats, and antibody fragments (ScFv, VHH, Fab).
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.
All projects are analyzed by A280 for antibody concentration and SDS-PAGE for purity assessment. Common optional QC add-ons include aSEC, endotoxin analysis, mass spectrometry services, and high-throughput ELISA. Additional analyses are available; please inquire for any custom QC requests.
Yes, we perform antibody affinity characterization by surface plasmon resonance (SPR) and bio-layer interferometry (BLI).
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.
Standard deliverables include: 2-5ug synthesized DNA construct (if performed by Azenta), antibodies provided as purified protein or supernatant, Certificate of Analysis (CoA).
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:
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, Azenta cannot guarantee final protein expression.
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 email@example.com.
CLONING AND MUTAGENESIS
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.
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.
Azenta currently only offers cloning vectors based on the pUC-GW-Kan/Amp backbone. We are unable to purchase proprietary vectors on your behalf.
If I cannot provide the starting material at the time of order confirmation, will the gene synthesis steps still be initiated?
Yes, the Project Management Team starts gene synthesis at the time of order confirmation.
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.
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.
For any information regarding on-going promotional discounts, please contact your sales representative.
What is the difference between site-directed mutagenesis, insert PCR-based mutagenesis, and TurboMUTANT?
|Site-Directed Mutagenesis||Insert PCR-Based Mutagenesis||TurboMUTANT|
|Plasmid Size Guidelines||10kb or less in size,
|> 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
Standard deliverables for Azenta 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
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.
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.
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.
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).
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).