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Dual primer emulsion PCR for next- generation DNA sequencing.

ABSTRACT: We have developed a highly sensitive single-molecule clonal amplification method called dual primer emulsion PCR (DPePCR) for next-generation DNA sequencing. The approach is similar in concept to standard emulsion PCR; however, in DPePCR both primers are attached to the beads, therefore following PCR amplification, both strands of the PCR products are attached to the beads. The ends of each strand can be freed for analysis by restriction digestion of the bridged PCR fragments, which allows efficient paired-end sequencing of fragment libraries.

SUBMITTER: Xu MY

PROVIDER: S-EPMC4049327 | biostudies-other | 2010 May

REPOSITORIES: biostudies-other

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Dual primer emulsion PCR for next- generation DNA sequencing.

Xu Ming Yan MY Aragon Anthony D AD Mascarenas Monica R MR Torrez-Martinez Norah N Edwards Jeremy S JS

BioTechniques 20100501 5

We have developed a highly sensitive single-molecule clonal amplification method called dual primer emulsion PCR (DPePCR) for next-generation DNA sequencing. The approach is similar in concept to standard emulsion PCR; however, in DPePCR both primers are attached to the beads, therefore following PCR amplification, both strands of the PCR products are attached to the beads. The ends of each strand can be freed for analysis by restriction digestion of the bridged PCR fragments, which allows effic ...[more]

PMID: 20569215

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