GENEWIZ Project Managers Are the Difference!

We have a track record of superior project management and a stringent quality system. GENEWIZ's experienced project managers are dedicated to ensuring your project's success. Our approach includes the assurance of milestones for each stage of the process, and clear communication to keep you well-informed.

• Dedicated Ph.D. project managers in New Jersey
• Strong molecular biology background
• Weekly project updates
• End-to-end support

Gene Synthesis Algorithms

Algorithms are the core of Gene Synthesis technology. The success of Gene Synthesis projects relies on two algorithms, the Gene Optimization Algorithm and the Oligo Design Algorithm. With the Gene Optimization Algorithm, a gene can be easily adapted to the codon frequency of almost any expression system, and optimized for protein expression. Advances in the Oligo Design Algorithm have led to greatly improved stability of Gene Synthesis technology.

Gene Optimization

Although the vast majority of organisms use the same genetic code (the standard genetic code), the usage frequency of certain codons can be very different from organism to organism (see the example below). This is also known as codon bias or codon usage.  Because of the difference in codon frequency, expression of a foreign gene in the expression host cells may result in reduced translational efficiency and low expression. In addition, variations to the standard genetic code, global and local GC content, inverted repeats and other mRNA secondary structures, undesirable motifs, including premature PolyA sites, may also negatively impact gene expression. The Gene Optimization Algorithm provides possible solutions for these potential problems.

Example: Arginine Codon Frequency Variation

The table below shows a comparison of arginine codon frequencies in Human, Drosophila, and E. coli (Grantham et al. 1986). Arginine is encoded by each of the six codons for various genes in the species.  The percentages indicate a preference (or bias) for an arginine codon in a particular species.  For example, E. coli prefers CGC (39%) and CGT (49%) to code for arginine.

Oligo Design

A Gene Synthesis project begins with oligo synthesis. An Oligo Design Algorithm can help design the oligos based on the full-length gene assembled. A good Oligo Design Algorithm can facilitate the design of oligos that meet the following criteria: within a certain length range with a relative uniform melting temperature; free of secondary structures; no mismatching or mis-hybridization.

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?

We clone the synthetic genes into the EcoRV site in pUC57 . A customer may also specify the restriction site for cloning.

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 construct 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 ug (depending on vector) of lyophilized plasmid DNA. To resuspend it, we recommend:

1. Spin the tube briefly to ensure that the contents are on the bottom of the tube
2. Resuspend the sample in a volume appropriate for your needs (such as 20-50 uL for ~0.25 to ~0.1 ug/uL) of either sterile, high purity water or TE (resuspension solution depends on your downstream applications; TE may interfere with various enzyme manipulations)
3. 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).