Library preparation and multiplex capture for massive parallel sequencing applications made efficient and easy.
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Neiman M, Sundling S, Grönberg H, Hall P, Czene K, Lindberg J, Klevebring D
PLoS ONE 7 (11) e48616 [2012-11-05; online 2012-11-05]
During the recent years, rapid development of sequencing technologies and a competitive market has enabled researchers to perform massive sequencing projects at a reasonable cost. As the price for the actual sequencing reactions drops, enabling more samples to be sequenced, the relative price for preparing libraries gets larger and the practical laboratory work becomes complex and tedious. We present a cost-effective strategy for simplified library preparation compatible with both whole genome- and targeted sequencing experiments. An optimized enzyme composition and reaction buffer reduces the number of required clean-up steps and allows for usage of bulk enzymes which makes the whole process cheap, efficient and simple. We also present a two-tagging strategy, which allows for multiplex sequencing of targeted regions. To prove our concept, we have prepared libraries for low-pass sequencing from 100 ng DNA, performed 2-, 4- and 8-plex exome capture and a 96-plex capture of a 500 kb region. In all samples we see a high concordance (>99.4%) of SNP calls when comparing to commercially available SNP-chip platforms.
PubMed 23139805
DOI 10.1371/journal.pone.0048616
Crossref 10.1371/journal.pone.0048616
pii: PONE-D-12-16041
pmc: PMC3489721