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High-fidelity PCR enzyme with DNA-binding domain facilitates de novo gene synthesis.

ABSTRACT: Nowadays enzymatic synthesis of genes is the most powerful tool for fast resolution of the various tasks in the field of basic and applied biological research. PCR-based gene assembly from overlapping oligonucleotides has become a widely used strategy. However, all the methods described in the literature are not perfect and need an extra processing step. In this study we are verifying Phusion high-fidelity polymerase as a tool to reduce nucleotide mismatches in de novo gene synthesis, thus facilitating subsequent cloning. To test the efficiency of the polymerase, we selected Fel d 4 gene, which is a 581 bp DNA sequence encoding the lipocalin allergen protein, one of the major cat allergens. The approach described here, therefore, would be useful in DNA sequences creation.

SUBMITTER: Dolgova AS

PROVIDER: S-EPMC5453909 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

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High-fidelity PCR enzyme with DNA-binding domain facilitates de novo gene synthesis.

Dolgova A S AS Stukolova O A OA

3 Biotech 20170601 2

Nowadays enzymatic synthesis of genes is the most powerful tool for fast resolution of the various tasks in the field of basic and applied biological research. PCR-based gene assembly from overlapping oligonucleotides has become a widely used strategy. However, all the methods described in the literature are not perfect and need an extra processing step. In this study we are verifying Phusion high-fidelity polymerase as a tool to reduce nucleotide mismatches in de novo gene synthesis, thus facil ...[more]

PMID: 28573398

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