GSM7230825
RNA was harvested using TRIzol method (Invitrogen) according to the manufacturer's instructions. miRNA sequencing and initial data analysis was performed by Arraystar Inc. (9700 Great Seneca Highway #171, Rockville, MD 20850). Total RNA of each sample was sequentially ligated to 3' and 5' small RNA adapters. cDNA was then synthesized and amplified using Illumina's proprietary RT primers and amplification primers. Subsequently, ~135-155 bp PCR amplified fragments were extracted and purified from the PAGE gel. And finally, the completed libraries were quantified by Agilent 2100 Bioanalyzer. The DNA fragments in the libraries were denatured with 0.1M NaOH to generate single-stranded DNA molecules, captured on Illumina flow cells, amplified in situ and finally sequenced for 36 cycles on Illumina HiSeq 2000 according to the manufacturer's instruction.