GSM2629457: C603; Bombyx mori; RNA-Seq
Total RNA was extracted using RNAiso Plus (TaKaRa, China) and dissolved in RNase-free water. The concentration of total RNA was determined using a NANODROP1000 microspectrophotometer (Thermo, USA) after treatment with DNase. The treated mRNA was enriched using oligo (dT) magnetic beads. By mixing with fragmentation buffer, the mRNA was fragmented into short fragments. Then, the first strand of cDNA was synthesized using random hexamer primers. Buffer, dNTPs, RNase H and DNA polymerase I were subsequently added to synthesize the second strand. Next, the double-stranded cDNA was purified with magnetic beads. End repair and 3’-end single A (adenine) nucleotide addition were then performed. Finally, sequencing adaptors were ligated to the fragments, and the fragments were enriched via PCR amplification. During the quality control (QC) step, an Agilent 2100 Bioanaylzer and an ABI StepOnePlus Real-Time PCR System were used to qualify and quantify the sample library. The library products were then ready for sequencing via the Illumina HiSeqTM 2000 platform at the Beijing Genomics Institute (BGI).
GEO
GSM2629457