A pyrosequencing-tailored nucleotide barcode design unveils opportunities for large-scale sample multiplexing.

ABSTRACT: Multiplexed high-throughput pyrosequencing is currently limited in complexity (number of samples sequenced in parallel), and in capacity (number of sequences obtained per sample). Physical-space segregation of the sequencing platform into a fixed number of channels allows limited multiplexing, but obscures available sequencing space. To overcome these limitations, we have devised a novel barcoding approach to allow for pooling and sequencing of DNA from independent samples, and to facilitate subsequent segregation of sequencing capacity. Forty-eight forward-reverse barcode pairs are described: each forward and each reverse barcode unique with respect to at least 4 nt positions. With improved read lengths of pyrosequencers, combinations of forward and reverse barcodes may be used to sequenc

SUBMITTER: Parameswaran P 

PROVIDER: S-EPMC2095802 | biostudies-literature | 2007

REPOSITORIES: biostudies-literature