Document Detail


Identification of genetic variants using bar-coded multiplexed sequencing.
MedLine Citation:
PMID:  18794863     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We developed a generalized framework for multiplexed resequencing of targeted human genome regions on the Illumina Genome Analyzer using degenerate indexed DNA bar codes ligated to fragmented DNA before sequencing. Using this method, we simultaneously sequenced the DNA of multiple HapMap individuals at several Encyclopedia of DNA Elements (ENCODE) regions. We then evaluated the use of Bayes factors for discovering and genotyping polymorphisms. For polymorphisms that were either previously identified within the Single Nucleotide Polymorphism database (dbSNP) or visually evident upon re-inspection of archived ENCODE traces, we observed a false positive rate of 11.3% using strict thresholds for predicting variants and 69.6% for lax thresholds. Conversely, false negative rates were 10.8-90.8%, with false negatives at stricter cut-offs occurring at lower coverage (<10 aligned reads). These results suggest that >90% of genetic variants are discoverable using multiplexed sequencing provided sufficient coverage at the polymorphic base.
Authors:
David W Craig; John V Pearson; Szabolcs Szelinger; Aswin Sekar; Margot Redman; Jason J Corneveaux; Traci L Pawlowski; Trisha Laub; Gary Nunn; Dietrich A Stephan; Nils Homer; Matthew J Huentelman
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2008-09-14
Journal Detail:
Title:  Nature methods     Volume:  5     ISSN:  1548-7105     ISO Abbreviation:  Nat. Methods     Publication Date:  2008 Oct 
Date Detail:
Created Date:  2008-09-30     Completed Date:  2008-10-21     Revised Date:  2012-04-26    
Medline Journal Info:
Nlm Unique ID:  101215604     Medline TA:  Nat Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  887-93     Citation Subset:  IM    
Affiliation:
The Translational Genomics Research Institute, Phoenix, Arizona 85004, USA. dcraig@tgen.org
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MeSH Terms
Descriptor/Qualifier:
Automatic Data Processing*
Genetic Variation*
Genome, Human*
Humans
Polymorphism, Single Nucleotide
Sequence Alignment
Grant Support
ID/Acronym/Agency:
R01 N5059873//PHS HHS; U01 HL086528/HL/NHLBI NIH HHS; U01 HL086528-03/HL/NHLBI NIH HHS
Comments/Corrections

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