Mutation discovery by targeted genomic enrichment of multiplexed barcoded samples.
Targeted genomic enrichment followed by next-generation DNA sequencing has dramatically increased efficiency of mutation-discovery efforts. We describe a protocol for genomic enrichment of pooled barcoded samples in a single assay that increases experimental flexibility and efficiency. We screened 770 genes (1.4 megabases) in thirty N-ethyl-N-nitrosourea (ENU)-mutagenized rats and identified known variants at >96% sensitivity as well as new mutations at a false positive rate < 1 in 8 megabases.
Hubrecht Institute, Developmental Biology and Stem Cell Research, Royal Netherlands Academy of Arts and Sciences and the University Medical Center Utrecht, Utrecht, The Netherlands.
Isaäc J
IJ
Nijman
Michal
M
Mokry
Ruben
R
van Boxtel
Pim
P
Toonen
Ewart
E
de Bruijn
Edwin
E
Cuppen
D000818:Animals; D001330:Electronic Data Processing; D005038:Ethylnitrosourea / Q000633:toxicity; D008297:Male; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D051381:Rats; D017422:Sequence Analysis, DNA
D005038:Ethylnitrosourea