Shotgun-sequencing of plant DNA: Selection og material and methods

Abstract

Initially, rbcLaand ITS2 were PCR amplified and Sanger sequenced for 564 species from 1900 specimens. To increase the amount of the genome covered, shotgun sequencing of 1670 species is in progress. Our Sanger analyses had a success rate of 74% and 85% for ITS2 and rbcLa, respectively. Overall success rate of Sanger sequencing decreased with later collection date in the year. Preliminary results of the shotgun sequencing and assembly of 277 specimens gives a success rate of 82% and 72% for rbcL and matK, respectively. Constructing a reference sequence library consisting of not only a few bar code loci, but full plastid and ribosomal sequences is beneficial as it provides a valuable resource for novel research avenues.