title: GSM3132936: FP2; Nelumbo nucifera; RNA-Seq
library_construction_protocol: Rhizomes of first-year planting  (FP) and consecutive planting  (CP) were harvested, flash frozen on dry ice, and RNA was harvested using Trizol reagent. 1 ug of total RNA was used for the construction of sequencing libraries by BGISEQ-500. Six rhizomes were randomly collected from both FP and CP groups, with three samples for each group. The total RNA of rhizomes was isolated by Trizol reagent, and the RNA quality was checked by an Agilent 2100 Bioanalyzer (Aglient Technologies, Santa Clara, CA, USA). Firstly, mRNA was enriched by oligo(dT) magnetic beads, and then interrupted to short fragments with the size of about 200 bp. The short fragments were used as templates to synthesize the double-strand cDNA by random primers. Additionally, sequencing adaptors were ligated to the fragments, and the required fragments were purified and amplified by PCR reaction. The library products were scanned for sequencing analysis via BGISEQ-500 (BGI, Wuhan, China).