title: GSM5100262: RNAs from seed produced at 23°C_germination II stage, ros_2; Arabidopsis thaliana; RNA-Seq
library_construction_protocol: Total RNAs were extracted from the frozen grinded tissues lots using the Macherey-Nagel NucleoSpin RNA Plus kit (Macherey-Nagel, GmbH & Co. KG, Germany) following manufacturer's instructions Samples were sent to BGI, Hong Kong, for library preparation. Libraries were constructed following a custom protocol from samples that passed quality controls (mass > 2µg, concentration>80ng/microl, OD260/280 ~= 2.00, OD260/230 ~= 2.20, RIN>6.5, 28S/18S<1.0, baseline smooth). After mRNA enrichment, RNA was fragmented and reverse transcribed to double-strand cDNA (dscDNA) by N6 random primer. The synthesized cDNA was subjected to end-repair and then was 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments. The ligation products were purified and many rounds of PCR amplification were performed to enrich the purified cDNA template using PCR primer, splint oligo and DNA ligase, followed by sequencing on BGISEQ-500 platform, generating an average 24M reads of 50bp per sample