Targeted Capture and High-Throughput Sequencing Using Molecular Inversion Probes (MIPs).
Molecular inversion probes (MIPs) in combination with massively parallel DNA sequencing represent a versatile, yet economical tool for targeted sequencing of genomic DNA. Several thousand genomic targets can be selectively captured using long oligonucleotides containing unique targeting arms and universal linkers. The ability to append sequencing adaptors and sample-specific barcodes allows large-scale pooling and subsequent high-throughput sequencing at relatively low cost per sample. Here, we describe a "wet bench" protocol detailing the capture and subsequent sequencing of >2000 genomic targets from 192 samples, representative of a single lane on the Illumina HiSeq 2000 platform.
Department of Genome Sciences, University of Washington School of Medicine, 3720 15th Ave NE, S413C, Box 355065, Seattle, WA, 98195-5065, USA. cantss@u.washington.edu.
Stuart
S
Cantsilieris
Department of Genome Sciences, University of Washington School of Medicine, 3720 15th Ave NE, S413C, Box 355065, Seattle, WA, 98195-5065, USA.
Holly A
HA
Stessman
Department of Genome Sciences, University of Washington School of Medicine, 3720 15th Ave NE, S413C, Box 355065, Seattle, WA, 98195-5065, USA.|Howard Hughes Medical Institute, University of Washington, Seattle, WA, 98195, USA.
Jay
J
Shendure
Department of Genome Sciences, University of Washington School of Medicine, 3720 15th Ave NE, S413C, Box 355065, Seattle, WA, 98195-5065, USA.|Howard Hughes Medical Institute, University of Washington, Seattle, WA, 98195, USA.
Evan E
EE
Eichler
D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D015335:Molecular Probes; D016133:Polymerase Chain Reaction
D015335:Molecular Probes
Exonuclease cleanup and gel electrophoresis; Massively parallel sequencing; Molecular inversion probes; Real-time PCR