GSM1933849: 6month; Mus musculus; miRNA-Seq
Total RNA was extracted from the Hippocampal tissue using Trizol Reagent (Invitrogen) according to the manufacturer’s instructions.Length of 18–30 nt small RNAs were isolated from total RNA using 15% denaturing polyacrylamide gel electrophoresis. The RNA isolated from each sample was used to construct sequencing libraries with the Illumina TruSeq Small RNA Sample Prep Kit, following the manufacturer's protocol. Briefly, 3′ and 5′ adapters were sequentially ligated to small RNA molecules and the obtained ligation products were subjected to a reverse transcription reaction to create single stranded cDNA. To selectively enrich those fragments that have adapter molecules on both ends, the cDNA was amplified with 15 PCR cycles using a common primer and a primer containing an index tag; this allows multiplexing and sequencing of different samples in a single lane of a flowcell. The amplified cDNA constructs were gel purified, and validated by checking the size, purity, and concentration of the amplicons on the Agilent Bioanalyzer. The libraries were pooled in equimolar amounts, and sequenced on an Illumina HiSeq 2000 instrument to generate 50 base reads. Image decomposition and quality values calculation were performed using the modules of the Illumina pipeline.
GEO Accession
GSM1933849