Characterization of de novo transcriptome for horseweed (Conyza canadensis) using GS-FLX 454 pyrosequencing
The cDNA sample was fractionated into smaller pieces (300–500 bp) that were subsequently polished (blunted). The short A and B adaptors were then ligated on to each resulting fragment, which provide priming sequences for both emulsion PCR amplification and pyrosequencing, forming the basis of the single-stranded template library. Approximately 15 µg of double-stranded cDNA was used for three sequencing runs using a Roche GS-FLX sequencer at the University of Illinois Keck Center using standard methods.