Decoding a substantial set of samples in parallel by massive sequencing
2011 (English)In: Plos One, ISSN 1932-6203, Vol. 6, no 3Article in journal (Refereed) Published
The dramatic increase of throughput seen in the eld of sequenceanalysis during the last years has opened up new possibilities of se-quencing a multitude of samples in parallel. Here we present a novelstrategy where the combination of two tags is used to link reads totheir origins in a pool of samples. The two tags are incorporated intwo steps leading to lowering of sample handling complexity by nearly100 times. The method described here enables accurate identi cationand typing of thousands of samples in parallel and is scalable. In thisstudy the system was designed to test 4992 samples using only 122 tags.
To proof the concept of two tagging method the highly polymor-phic 2nd exon of DLA-DRB1 in dogs and wolves was sequenced usingthe 454 GS FLX Titanium Chemistry. By requiring a minimum se-quence depth of 20 reads per sample, 94% of the successfully ampli edsamples were genotyped. In addition, the method allowed digital de-tection of chimeric fragments. These results demonstrate that it ispossible to sequence thousands of samples in parallel without complexpooling patterns or primer combinations. Furthermore, the method isscalable and increasing the sample size by 960 samples requires only10 additional tags.
Place, publisher, year, edition, pages
San Fransisco: PUBLIC LIBRARY SCIENCE , 2011. Vol. 6, no 3
HIGH-THROUGHPUT; HIGH-RESOLUTION; DNA; PROBES
IdentifiersURN: urn:nbn:se:kth:diva-24353DOI: 10.1371/journal.pone.0017785ISI: 000288170900047ScopusID: 2-s2.0-79952430921OAI: oai:DiVA.org:kth-24353DiVA: diva2:347893
FunderScience for Life Laboratory - a national resource center for high-throughput molecular bioscience
QC 20100906 Uppdaterad från manuskript till artikel i tidskrift 201104072010-09-062010-09-032012-11-16Bibliographically approved