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Frequently Asked Questions
What lengths of synthetic genes can
you synthesize?
We can synthesize gene sequences from 100 bp to 50 kb, using
short DNA oligos as building blocks.
Can you synthesize sequences with
high GC content or repeats?
Yes. GENEWIZ's proprietary technologies enable us to synthesize
sequences with difficult stretches. Our bioinformatics platform can optimize
your gene for synthesis. We carefully review every sequence, and provide a
customized quote including the turnaround time and price.
How do you synthesize genes?
Single-stranded oligonucleotides are designed, synthesized, and
assembled using different protocols depending on the sequence characteristics.
The assembled full-length gene is subsequently cloned into the vector
pUC57, and the insert is verified by DNA
sequencing and restriction digestion.
Is there any extra charge in addition
to the per bp price?
We provide a customized quote for every sequence. For typical
gene sequences (i.e. less than 2kb, free of difficult stretches like highly
repetitive, AT-rich, or GC-rich fragments) that do not require subcloning into
the customer's own vector, there will be no extra charge to the per bp price.
For difficult sequences, or projects with subcloning requirements, all
additional charges will be shown on the customized quote. Your price will be
exactly as quoted.
What restriction sites do you use to
clone the synthetic genes into pUC57?
pUC57 contains several common restriction sites.
Standard cloning into pUC57 typically occurs via EcoRV,
however the exact restriction site(s) used for each sequence may vary;
specific restriction sites within the MCS may be maintained or excluded upon request.
What antibiotic resistance gene is
found in pUC57?
Ampicillin
Can you clone the synthetic gene
directly into my vector and save the step of cloning into pUC57?
We are happy to clone your sequence into your vector. Cloning
into
pUC57 is part of our standard procedure for gene synthesis, helping to
ensure the accurate construction of your sequence. As part of our quality
control, all constructs are cloned and sequence verified in pUC57 prior to
custom cloning into your vector.
My vector is a commonly used
expression vector. Do you have it in stock?
Vectors other than
pUC57 must be supplied by the customer. If you need the synthetic gene
cloned into your own vector, please send GENEWIZ the vector DNA and pertinent
information.
How can I be assured that I'm getting
the correct sequence?
We have controls in place throughout a project to ensure the
accurate synthesis of your sequence. First, the exact sequence to be synthesized
is included in the custom quote for your project, facilitating your review and
confirmation prior to project initiation. Second, sequence verification during
the assembly process helps confirm that any subassemblies contain the intended
sequence. Third, the final mini-prep is clonally isolated and confirmed to match
the quoted sequence. Finally, the electronic data package delivered in parallel
with your construct contains the original sequencing data files from the
construct verification for your independent review.
Also included in this package are: a certificate of
authentication with a restriction digest image of the sample, an alignment of
the sequencing data with the complete insert sequence, and the predicted
complete vector sequence.
What is your recommended DNA
resuspension protocol?
The sample tube from GENEWIZ contains 2-5 µg (depending on
vector) of lyophilized plasmid DNA. To resuspend it, we recommend:
- Spin the tube briefly to ensure that the contents are on the
bottom of the tube
- Resuspend the sample in a volume appropriate for your needs (such
as 20-50 µL for ~0.25 to ~0.1 µg/µL) of either sterile, high purity water or TE
(resuspension solution depends on your downstream applications; TE may interfere
with various enzyme manipulations)
- Vortex briefly, let sit 2-10 minutes on ice, then vortex again.
Do you have any mechanism for
addressing potential biosecurity concerns that may arise with the genes that you
synthesize?
GENEWIZ is fully aware that Gene Synthesis technology
potentially enables the de novo reconstruction of dangerous pathogens,
and of the need for an in-house mechanism to safeguard against intentional or
unintentional abuse of the genes that we synthesize. We are monitoring the
progress of the Screening Framework Guidance for Synthetic Double-Stranded DNA
Providers, drafted by U.S. Department of Health and Human Services (HHS).
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