title: GSM3035338: 2cav; Homo sapiens; miRNA-Seq
library_construction_protocol: mirVana miRNA isolation Kit (AM1560; Ambion) was used to isolate total RNA containing small RNA from the tissue specimens. The eluted RNA underwent DNase I treatment in solution for 10 min at RT by adding 10 µl of buffer RDD and 2.5 µl of DNase I, both extracted from RNeasy Micro Kit (74004; Qiagen). Thereafter, 10 µl of 5 M NaCl and 152 µl of 100% Ethanol were added and were thoroughly mixed followed by loading into the RNeasy column for end point elution. Total RNA was then eluted with 14 µl of RNase-free water. Quantification and quality of total RNA was carried out with the NanoDrop ND-1000 spectrophotometer (Peqlab). RNA integrity was detected with an Agilent Bioanalyzer 2100 using the RNA 6000 Nano Assay (5067-1511; Agilent Technologies). Only RNA extracts with a RNA integrity number (RIN) values ≥ 7 underwent further deep sequencing analysis. A starting material of 50-100 ng total RNA was used to generate RNA sequencing library using the TruSeq® Small RNA Library Prep Kits v2 (Illumina). The libraries were sequenced on Illumina HiSeq2500 using TruSeq SBS Kit v3-HS (50 cycles, single ended run) with an average of 10 x106 reads per RNA sample.
attributes.tag: GEO Accession
attributes.value: GSM3035338