GSM2061358: HeLa; Scrambled siRNA; Homo sapiens; ncRNA-Seq
Total cellular RNA was extracted with RNeasy Mini kit (Qiagen) in accordance with the manufacturer's instructions, prior to sequencing. Ribosomal RNAs (rRNA) were removed from total RNA samples using Ribo-Zero™ rRNA Removal Kits (Epicentre). The total RNA samples (after depletion of rRNA) were size-fractionated on a 6% TBE-Urea polyacrylamide gel to enrich for molecules in the range of 50-400nt. Small RNA libraries and sequencing for each sample were performed by LC Sciences (Houston, TX). In brief, RNA fragments were dephosphorylated and ligated with 3′ RNA adaptor. The resulting ligated RNA fragments were reversed-transcribed and ligated with 5′ DNA/DNA adaptor and PCR amplified (15 cycles) using primers on both adaptors. Subsequently, PCR products were purified and small RNA library was sequenced on Illumina GAIIx (Illumina, Inc., Santa Clara, CA) for single-end 35-mers or on Illumina HiSeq for single-end 51-mers following the manufacturer’s protocol. Raw sequence FASTQ files were generated by using Illumina’s software pipeline..
GEO
GSM2061358
GEO Accession
GSM2061358