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Reflex: intramolecular barcoding of long-range PCR products for sequencing multiple pooled DNAs.

ABSTRACT: We present an intramolecular reaction, Reflex™, to derive shorter, sequencer-ready, daughter polymerase chain reaction products from a pooled population of barcoded long-range polymerase chain reaction products, whilst still preserving the cognate DNA barcodes. Our Reflex workflow needs only a small number of primer extension steps to rapidly enable uniform sequence coverage of long contiguous sequence targets in large numbers of samples at low cost on desktop next-generation sequencers.

SUBMITTER: Casbon JA 

PROVIDER: S-EPMC3664814 | biostudies-other | 2013 May

REPOSITORIES: biostudies-other

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Reflex: intramolecular barcoding of long-range PCR products for sequencing multiple pooled DNAs.

Casbon James A JA   Slatter Andrew F AF   Musgrave-Brown Esther E   Osborne Robert J RJ   Lichtenstein Conrad P CP   Brenner Sydney S  

Nucleic acids research 20130410 10

We present an intramolecular reaction, Reflex™, to derive shorter, sequencer-ready, daughter polymerase chain reaction products from a pooled population of barcoded long-range polymerase chain reaction products, whilst still preserving the cognate DNA barcodes. Our Reflex workflow needs only a small number of primer extension steps to rapidly enable uniform sequence coverage of long contiguous sequence targets in large numbers of samples at low cost on desktop next-generation sequencers. ...[more]

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