GSM4192150: T2-1; Pinellia ternata; RNA-Seq
Five seedlings collected from each group were mixed and thoroughly ground to powder  with liquid nitrogen, and approximately 50 mg per pooled sample was prepared to use. Total RNA was isolated from each replicated sample according to the instructions of OmniPlant RNA Kit  (CWBIO, Beijing, China) We constructed the cDNA library using NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, USA); brief steps are described below. Firstly, the mRNA was enriched by oligo (dT)-attached magnetic beads and fragmented into short pieces, which were taken as templates for the first-strand and second-strand cDNA synthesis.  Then, the remaining overhangs into blunt ends were converted by ligase and polymerase. The resulting fragments were end-repaired by inserting an “A” base to the 3' ends of the cDNA. And the NEBNext adapters with a hairpin loop structure were ligated to fragments. To select suitable cDNA fragments, Agencourt AMPure XP 60ml Kit (Beckman Coulter, USA) was applied to purify the library fragments. Next, the products amplified by polymerase chain reaction (PCR), which were used to create sequencing libraries.  And 350-450 bp fragments were recovered using QIAquick Gel Extraction Kit (QIAGEN, Germany). It's crucial of quality testing for the library to ensure that the library meets the requirements of sequencing
GEO Accession
GSM4192150