Anemone flaccida root transcriptome
Five microliters of PCR products were detected by electrophoresis on 1% agarose gels to determine the amplification efficiency and quality. The amplified ds cDNA were purified using the PureLink PCR Purification Kit (Life Technologies, USA). The purified cDNA was then digested with BsgI (New England Biolabs, USA) overnight at 37℃ and recovered with the QIAquick PCR Purification Kit (Qiagen, Germany). Finally, about 10 μg of ds cDNAs was used for pyrosequencing with the GS FLX Titanium Rapid Library Preparation Kit (Roche, Germany) following the manufacturer´s instructions.