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Dual primer emulsion PCR for next- generation DNA sequencing.
Biotechniques. 2010 May;48(5):409-12
Authors: Xu MY, Aragon A, Mascarenas M, Torrez-Martinez N, Edwards J
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. Address correspondence to Jeremy S. Edwards, University of New Mexico, Molecular Genetics and Microbiology, 915 Camino de Salud NE MSC08 4660, Albuquerque, NM 87131, USA. e-mail:
PMID: 20569215 [PubMed - in process]
More...
Dual primer emulsion PCR for next- generation DNA sequencing.
Biotechniques. 2010 May;48(5):409-12
Authors: Xu MY, Aragon A, Mascarenas M, Torrez-Martinez N, Edwards J
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. Address correspondence to Jeremy S. Edwards, University of New Mexico, Molecular Genetics and Microbiology, 915 Camino de Salud NE MSC08 4660, Albuquerque, NM 87131, USA. e-mail:
PMID: 20569215 [PubMed - in process]
More...