Targeted exon sequencing by in-solution hybrid selection.
This unit describes a protocol for the targeted enrichment of exons from randomly sheared genomic DNA libraries using an in-solution hybrid selection approach for sequencing on an Illumina Genome Analyzer II. The steps for designing and ordering a hybrid selection oligo pool are reviewed, as are critical steps for performing the preparation and hybrid selection of an Illumina paired-end library. Critical parameters, performance metrics, and analysis workflow are discussed.
Blumenstiel
Brendan
B
Broad Institute, Cambridge, Massachusetts, USA.
Cibulskis
Kristian
K
Fisher
Sheila
S
DeFelice
Matthew
M
Barry
Andrew
A
Fennell
Tim
T
Abreu
Justin
J
Minie
Brian
B
Costello
Maura
M
Young
Geneva
G
Maquire
Jared
J
Kernytsky
Andrew
A
Melnikov
Alexandre
A
Rogov
Peter
P
Gnirke
Andreas
A
Gabriel
Stacey
S
D005091:Exons / Q000235:genetics*; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*; D012996:Solutions
D012996:Solutions