GSM3223386: Si rep2; Cucumis sativus; RNA-Seq
Total RNA was harvested using Trizol reagent following manufacture`s protocol (Roche, Mannheim, Germany). RNA-Seq library construction and sequencing were conducted by Gene Denovo Co. (Guangzhou, China). The poly(A)-containing mRNA molecules were purified from 10 mg of total RNA using poly-T oligo-attached magnetic beads. The extracted mRNAs were fragmented into 200-bp-long pieces using RNA Fragmentation Buffer (Kapa, Biosystems). The cleaved Illumina Sequencing mRNA fragments were reverse-transcripted into first-strand cDNA using random primers, followed by second-strand cDNA synthesis using DNA Polymerase I and RNase H. Then, the cDNA fragments went through an end repair process, the addition of a single “A” base, and ligation of the adapter sequences. Finally, the cDNA products were purified and enriched to construct libraries for non-strand-specific RNA-Seq.
GEO Accession
GSM3223386