Custom synthetic genes
Each gene is manufactured by total chemical synthesis starting with oligonucleotides.
The synthetic DNA is built and cloned into our standard pUC-derived plasmid or any vector of your choice. Transformed colonies are screened, a first set of five positive clones are selected, amplified and purified. Agarose gel electrophoresis run allows for size and flanking restriction site verification. Your gene is then sequenced in both orientations and the fluorescence traces files are provided so you may verify sequence correctness. We guarantee 100% accurate sequence.
- Fig 1. Every positive clone undergoes stringent quality control procedures including restriction site analysis and thorough coverage by double-strand DNA sequencing.
- Click on figure to enlarge
Standard synthesis service comprises
- Codon optimization for heterologous expression: Optimize sequences towards a more favorable codon assortment and thus improving expression in your host.
- Sequence design: Addition or removal of sequence features including N or C purification tags, removal of restriction sites, amino acid changes for structure activities studies, etc.
- Total de novo gene synthesis to your specifications.
- Cloning, screening, amplification and purification of your clone: our standard procedure includes cloning into a pUC-derived plasmid. Optionally, we would clone your gene into any vector that you would provide.
- Complete double-strand sequencing of your gene
Deliverables include:
5 µg of purified plasmid DNA, sequencing data and full synthesis report.
Site directed mutagenesis
With this service, we would introduce any types of mutation into your gene of interest, including deletions, insertions, or substitutions.
Single or multiple changes in a single gene may be achieved.
Deliverables include 5 µg of purified plasmid DNA and double-stand sequencing data.
- Fig 2. Site directed mutagenesis, single base-pair substitution.
- Click on figure to enlarge
Additional services:
- Plasmid preparation
- Subcloning services
