Description: NZYGene Synthesis kit was designed to easily synthesise any DNA fragment using PCR assembly & amplifications protocols. The kit includes an additional error correction step with UltraPrecise T7 Endonuclease I, which is a new generation of enzymes that recognizes and cleave DNA sequences with DNA mismatches. The integrated NZYGene Synthesis workflow follows simple, rapid and accurate protocols to facilitate whole gene synthesis. Briefly, the NZYGene synthesis system starts with the design of overlapping oligonucleotides required to produce the desired genes. Overlapping oligonucleotides are assembled into a new DNA fragment by PCR using a high-fidelity DNA polymerase. The innovative UltraPrecise T7 Endonuclease I enzyme is added to recognise and cleave DNA mismatches resulting from gene assembly allowing the removal of errors that accumulate during the PCR reaction. A final amplification is required to produce error-corrected DNA fragments that are directly cloned into pHTP0 cloning vector following the NZYEasy cloning system. No further additional steps for the removal of errors from synthesised genes, e.g. site-directed mutagenesis, are required. All protocols of this kit were optimized to display high efficiencies and a reduced error rate associated with the production of synthetic genes.
This kit has been successfully used in high-throughput (HTP) platforms for the efficient gene synthesis of a large number of genes at a scale compatible with the functional screen of hundreds to thousands of genes.
– In vitro gene synthesis
– GS DNA Polymerase (1U/ μL)
– Reaction Buffer for GS DNA Polymerase (10x)
– dNTP mix (2 mM)
– MgSO4 (25 mM)
– UltraPrecise T7 Endonuclease I
– Reaction Buffer for UltraPrecise T7 Endonuclease I (10x)
– pHTP0 vector
– Reaction Buffer (10x)
– NZYEasy enzyme mix
– Positive control
– NZYStar Competent Cells
– Competent Cells Control Plasmid (0.1 ng/µL)