[metagenomics-jclub] May 11th metagenomics J-club: Bait and Catch!

[Fan]

Hi all,

I will be presenting the paper on Bait and Catch techniques for the next 
metagenomics journal club (May 11th, 2011, next Wednesday). The abstract 
is stated beneath and attached find the paper.

See you next week!

Cheers!

Fan
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Targeting genomic loci by massively parallel sequencing requires new 
methods to enrich templates to be sequenced. We
developed a capture method that uses biotinylated RNA ‘baits’ to fish 
targets out of a ‘pond’ of DNA fragments. The RNA is
transcribed from PCR-amplified oligodeoxynucleotides originally 
synthesized on a microarray, generating sufficient bait for
multiple captures at concentrations high enough to drive the 
hybridization. We tested this method with 170-mer baits that target
415,000 coding exons (2.5 Mb) and four regions (1.7 Mb total) using 
Illumina sequencing as read-out. About 90% of uniquely
aligning bases fell on or near bait sequence; up to 50% lay on exons 
proper. The uniformity was such that B60% of target
bases in the exonic ‘catch’, and B80% in the regional catch, had at 
least half the mean coverage. One lane of Illumina
sequence was sufficient to call high-confidence genotypes for 89% of the 
targeted exon space.
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