Genomic reduction assisted single nucleotide polymorphism discovery using 454-pyrosequencing

Peter J Maughan; Joshua A Udall; Eric N Jellen

doi:10.1007/978-1-4939-1966-6_13

Genomic reduction assisted single nucleotide polymorphism discovery using 454-pyrosequencing

Methods Mol Biol. 2015:1245:169-82. doi: 10.1007/978-1-4939-1966-6_13.

Authors

Peter J Maughan¹, Joshua A Udall, Eric N Jellen

Affiliation

¹ 5144 LSB, Department of Plant and Wildlife Sciences, Brigham Young University, 150 East Bulldog Boulevard, Provo, UT, 84602, USA, Jeff_Maughan@byu.edu.

PMID: 25373757
DOI: 10.1007/978-1-4939-1966-6_13

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biotin / metabolism
Chromatography, Gel
DNA Barcoding, Taxonomic
Genomics / methods*
Polymerase Chain Reaction
Polymorphism, Single Nucleotide / genetics*
Restriction Mapping
Sequence Analysis, DNA / methods*
Streptavidin / metabolism

Substances

Biotin
Streptavidin