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2013 (engelsk)Inngår i: Scientific Reports, E-ISSN 2045-2322, Vol. 3, s. 1186-Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]
Here we demonstrate the use of short-read massive sequencing systems to in effect achieve longer read lengths through hierarchical molecular tagging. We show how indexed and PCR-amplified targeted libraries are degraded, sub-sampled and arrested at timed intervals to achieve pools of differing average length, each of which is indexed with a new tag. By this process, indices of sample origin, molecular origin, and degree of degradation is incorporated in order to achieve a nested hierarchical structure, later to be utilized in the data processing to order the reads over a longer distance than the sequencing system originally allows. With this protocol we show how continuous regions beyond 3000 bp can be decoded by an Illumina sequencing system, and we illustrate the potential applications by calling variants of the lambda genome, analysing TP53 in cancer cell lines, and targeting a variable canine mitochondrial region.
Emneord
Human Genome, Structural Variation, Domestic Dog, Dna, Mutations
HSV kategori
Identifikatorer
urn:nbn:se:kth:diva-105124 (URN)10.1038/srep01186 (DOI)000315767100001 ()23470464 (PubMedID)2-s2.0-84875360897 (Scopus ID)
Forskningsfinansiär
EU, FP7, Seventh Framework Programme, 222913Swedish Research Council
Merknad
QC 20130405. Updated from submitted to published.
2012-11-162012-11-162024-03-15bibliografisk kontrollert