GenTitan Gene Fragments

If a sequence doesn’t pass our GenTitan Gene Fragment difficulty screening filter, we recommend using our GenSmart codon optimization tool to optimize the sequences. If the optimization isn’t successful, our US gene synthesis service is available for clonal products.

To enhance the likelihood of successful synthesis:

• Keep repeat length below 20 base pairs.
• Avoid homopolymer (e.g., poly-A or poly-T) lengths longer than 15 base pairs.
• Keep the GC% content between 25-65% within 100 bp window. Using our GenSmart codon optimization tool can help improve the GC% content.

We guarantee 100% delivery of the qualified sequences. In the rare event of synthesis failure, we will provide clonal genes to ensure your research continues smoothly.

GenTitan gene fragments are not verified by NGS or Sanger sequencing due to the low cost and fast turnaround time requirements. However, due to the low error rate of our synthesis process, sequencing approximately 2-3 colonies is estimated to obtain a sequence-perfect clone with 95% confidence. If sequence-verified genes are required, please consider our US gene synthesis service.

Yes, gene fragments are purified using a bead-based method. Occasionally, there may be carryover of smaller DNA fragments.

Adapters are universal sequences added to each end of the gene fragment during synthesis. Fragments with adapters are compatible with multiple assembly methods, including restriction digest/ligation, Gateway, Golden Gate, and TOPO cloning. Below is the detailed information about the adapter.

The QC process for GenTitan Gene Fragments involves gel electrophoresis to check DNA size and purity, as well as UV reading for concentration.

Gene fragments are typically delivered in a freeze-dried format in 96-well plates. Options for 2ml microcentrifuge tubes or 384-well plates are also available, with additional charges or turnaround time (TAT) for specific plate types (e.g., Echo plate). Gene fragments can also be delivered in LTE buffer (10 mM Tris-HCl, pH 7.5; 0.1 mM EDTA), with no additional shipping charge. The concentration of gene fragments can be normalized to 20ng/µl with no extra charges.

• Dried DNA fragments are shipped at room temperature. Upon receipt of your gene fragments, the following steps are recommended: Briefly centrifuge the tube/plate to pellet the material to the bottom prior to opening.
  Add 1x TE (10 mM Tris, 0.1 mM EDTA) to the tube/plate for your desired final concentration, briefly vortex and centrifuge. We do not recommend resuspension in water. A minimum 10 ng/µl concentration is recommended for a stock solution.
• For long term storage, freeze gene fragments at -20°C or even better at -80°C and avoid repeated freeze-thaw cycles by preparing aliquots of the stock.

The default deliverable is 500ng (normalized) per fragment.

For restriction cloning, customer can design restriction sites within the adapters. The cloning process is similar to working with PCR products.

We do not recommend PCR amplification of the products after receipt. Gene fragments are purified using a bead-based method, which may result in the carryover of smaller DNA fragments. These smaller fragments tend to amplify more efficiently during PCR, leading to smears or additional smaller bands.

Typically, sequencing approximately 2-3 colonies should yield a sequence-perfect clone with 95% confidence. In some cases, due to the nature of DNA sequences, additional colony screenings may be necessary. When using two or more gene fragments for assembly and cloning, we strongly recommend screening more colonies or first confirming the correct individual gene fragment before proceeding with the assembly.

Currently our GenTitan Gene Fragment synthesis does not support N and K mixed bases. Please check our DNA Mutant Library for NNK library construction.

• Lyophilized format: Store at -20°C or -80°C for up to 2 years, or at room temperature for up to 6 months.
• Low TE buffer: Store at 4°C for up to 6-12 months.
• For long-term storage, keep at -20°C or -80°C and aliquot to minimize freeze-thaw cycles.