title: GSM9539427: Chlamydomonas reinhardtii WT, 24 h darkness, biological replicate 3; Chlamydomonas reinhardtii; RNA-Seq
library_construction_protocol: Total RNA was extracted using the RNAprep Pure Plant Plus Kit (TIANGEN, Cat. no. DP441) and treated with RNase-Free DNase I. Fragmentation was carried out using divalent cations under elevated temperature in First Strand Synthesis Reaction Buffer(5X). First strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase(RNase H-). Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. Remaining overhangs were converted into blunt ends via exonuclease/polymerase activities. After adenylation of 3' ends of DNA fragments, Adaptor with hairpin loop structure were ligated to prepare for hybridization. In order to select cDNA fragments of preferentially 370~420 bp in length, the library fragments were purified with AMPure XP system.