Genomic reduction assisted single nucleotide polymorphism discovery using 454-pyrosequencing.
We report the development of a simple genomic reduction protocol based on 454-pyrosequencing technology that discovers large numbers of single nucleotide polymorphisms (SNP) from pooled DNA samples. The method is based on the conservation of restriction endonuclease sites across samples and biotin separation for genomic reduction and the addition of multiplex identifier (MID) barcodes to each of the pooled samples to allow for postsequencing deconvolution of the pooled DNA fragments and SNP discovery.
5144 LSB, Department of Plant and Wildlife Sciences, Brigham Young University, 150 East Bulldog Boulevard, Provo, UT, 84602, USA, Jeff_Maughan@byu.edu.
Peter J
PJ
Maughan
Joshua A
JA
Udall
Eric N
EN
Jellen
D001710:Biotin / Q000378:metabolism; D002850:Chromatography, Gel; D058893:DNA Barcoding, Taxonomic; D023281:Genomics / Q000379:methods*; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA / Q000379:methods*; D019809:Streptavidin / Q000378:metabolism
D001710:Biotin; D019809:Streptavidin