Journal article

High-quality DNA sequence capture of 524 disease candidate genes

Peidong Shen, Wenyi Wang, Sujatha Krishnakumar, Curtis Palm, Aung-Kyaw Chi, Gregory M Enns, Ronald W Davis, Terence P Speed, Michael N Mindrinos, Curt Scharfe

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | NATL ACAD SCIENCES | Published : 2011

Abstract

The accurate and complete selection of candidate genomic regions from a DNA sample before sequencing is critical in molecular diagnostics. Several recently developed technologies await substantial improvements in performance, cost, and multiplex sample processing. Here we present the utility of long padlock probes (LPPs) for targeted exon capture followed by array-based sequencing. We found that on average 92% of 5,471 exons from 524 nuclear-encoded mitochondrial genes were successfully amplified from genomic DNA from 63 individuals. Only 144 exons did not amplify in any sample due to high GC content. One LPP was sufficient to capture sequences from <100-500 bp in length and only a single-tu..

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University of Melbourne Researchers

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Awarded by National Eye Institute



Awarded by NATIONAL CANCER INSTITUTE


Awarded by NATIONAL EYE INSTITUTE


Funding Acknowledgements

We thank Mamta Maheshwary and Keyi Liu for technical assistance and Rita Horvath, Kate Rauen, and Iris Schrijver for providing DNA samples. This work was supported by Grant R01 EY016240 from the National Eye Institute (to R.W.D. and C.S.) and Grants P30 CA016672 and 3U24CA143883 (to W.W.).