455 sequencing of Digitalis purpurea transcriptome
The single-stranded cDNA was prepared from 2 *g total RNA of Digitalis purpurea, using the SMART cDNA Synthesis kit and the modified poly(T) primer (5'-AAGCAGTGGTATCAACGCAGA GTGCAG T(20)VN-3') The double-stranded cDNA was amplified by PCR polymerase Advantage II. After removing those less than 300 bp using the PureLinkTM PCR Purification kit (Invitrogen), PCR products were digested with BsgI at 37 ? for 8 hours and then purified using the QIAquick PCR Purification kit (Qiagen). Pyrosequencing was performed using the 454 GS FLX Titanium Platform.