1. Front Cover.cdr - CORE

Session 2. Systems biology and -omics
A B S T R A C T B O O K – A B S T R A C T S O F T A L K S
RAD SEQUENCING OF PLANTS
Justin Pachebat
IBERS, Aberystwyth University, Aberystwyth, United Kingdom
E-mail: jip@aber.ac.uk
RAD sequencing is a form of reduced representation sequencing that utilises the highthroughput
illumina sequencing platform to generate SNPs for genotyping large numbers
of individuals in a population. The procedure involves digestion of genomic DNA with a
rare cutting restriction enzyme, ligation of a barcoded illumina compatible sequencing
adapter to the digested DNA, followed by pooling of samples, sonication to frgament the
DNA, and subsequent ligation of a second sequencing adapter and PCR amplification of
the multiplex library. These multiplexed samples are sequenced on an Illumina platform
and reads assigned to individual plants via the barcodes, and SNPs identified in the reads.
Here we describe how RAD sequencing can be applied to plants, such as Lolium perenne
for SNP genotyping, despite the lack of a reference genome.
20
X X I V S P P S C O N G R E S S 2 0 1 1