Poolde RNA of Corydalis Bulbs
Developing bulbs were harvested separately at Day0, 10 and 30, frozen immediately in liquid nitrogen and stored at -80℃ till use. Total RNA was extracted using TRIplant reagent (BioTeke, Beijing, China) following the manufacturer’s instructions, quantified spectrophotometrically at 260 nm. The equal quantities of high-quality total RNA from each of the above three stages of bulbs were pooled and submitted to Beijing Genomics Institute (BGI, Shenzhen, China) for Illumina paired-end cDNA library preparation and HiSeq™ 2000 sequencing.     mRNA was purified from 20 μg of DNaseI-treated total RNA with a RIN of ≥8 using oligo(dT) magnetic beads and then interrupted into short fragments by heat treatment in the fragmentation buffer. The cleaved mRNA fragments were reverse transcribed into the first cDNA strands using random hexamer primers followed by the second-strand cDNA synthesis using DNA polymerase I and RNase H. The dscDNAs were purified using a QiaQuick PCR extraction kit (Qigen), washed with the elution buffer and proceeded with end repair, Poly(A) tailing addition and sequencing adapter ligation. The ligated products were run on the agarose gel and the suitable fragments were size-selected for PCR amplification as the templates. The PCR libraries were purified, quantified and qualified via Agilent 2100 Bioanalyzer and ABI StepOnePlus Real-Time PCR System and sequenced using Illumina HiSeq™ 2000.