A new implementation of high-throughput five-dimensional clone pooling strategy for BAC library screening.

ABSTRACT:

Background

A five-dimensional (5-D) clone pooling strategy for screening of bacterial artificial chromosome (BAC) clones with molecular markers utilizing highly-parallel Illumina GoldenGate assays and PCR facilitates high-throughput BAC clone and BAC contig anchoring on a genetic map. However, this strategy occasionally needs manual PCR to deconvolute pools and identify truly positive clones.

Results

A new implementation is reported here for our previously reported clone pooling strategy. Row and column pools of BAC clones are divided into sub-pools with 1~ 2 x genome coverage. All BAC pools are screened with Illumina's GoldenGate assay and the BAC pools are deconvoluted to identify individual positive clones. Putative positive BAC clones are then further analyzed to find po

SUBMITTER: You FM 

PROVIDER: S-EPMC3016418 | biostudies-literature | 2010 Dec

REPOSITORIES: biostudies-literature