Meeting Abstract
118.7 Tuesday, Jan. 7 11:45 RADseq: Minimizing the cost of population genomics BIRD, C E*; FURINESS, S; HOGAN, J D ; HOBI LAB, ; Texas A&M Univ. – Corpus Christi chris.bird@tamucc.edu
Restriction site associated DNA sequencing (RADseq) have enabled population genomic research on non-model species, but the original protocols from the University of Oregon are quite expensive to perform. In order to address high costs, strategies were developed that involve using restriction enzymes to create fragments of a particular size or range of sizes. These size selection strategies, while effective for high quality, high molecular weight DNA, are not suited to degraded DNA where fragment size is not indicative of identity. Consequently, it is extremely difficult to achieve substantial coverage of targeted loci with most intermediate quality DNA samples that were isolated for mtDNA and microsatellite analysis. Here, we present modifications to the Oregon RADseq protocol that dramatically reduce costs to the same level as digest-size select-sequence methods, while maintaining its superiority in processing degraded samples. Our method, termed RADseq, requires only small amounts (10-100 ng) of partially degraded DNA in order to produce libraries for Illumina sequencing. RADseq facilitates the population genomic analysis of a wide range of samples at the minimal cost.
