GSM2751773: NKL; Musa acuminata; RNA-Seq
The improved CTAB (cetyltrimethylammonium bromide) method was used for RNA extraction (Elizabeth et al. 2008) Total RNA samples were first treated with DNase I to degrade any possible DNA contamination. The mRNA was enriched using oligo(dT) magnetic beads. After mixing with fragmentation buffer, the mRNA was fragmented into short fragments. The first-strand cDNA was synthesized using a random hexamer primer. Buffer, dNTPs, RNase H, and DNA polymerase I were then added to synthesize the second-strand cDNA. The double-stranded cDNA was purified using magnetic beads. End repair and 3'-end single nucleotide A (adenine) addition were then performed. Finally, sequencing adaptors were ligated to the fragments and the fragments were enriched by polymerase chain reaction (PCR) amplification. During the quality control (QC) step, a 2100 Bioanalyzer (Agilent, Santa Clara, CA, USA) and a StepOnePlus Real-Time PCR System (Applied Biosystems Inc., Foster City, CA, USA) were used to qualify and quantify sequences for the sample library
GEO Accession
GSM2751773