GSM1363233: sRNA of GC-1spg; Mus musculus; ncRNA-Seq
Total RNA was extracted from GC-1spg and GC-2spd (ts) cell lines by Trizol (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. The RNA samples were incubated with 10 U DNaseI (Takara, Dalian, China) for 30 min at 37°C to remove residual genomic DNA. The quality and quantity of the purified RNA were determined by measuring the absorbance at 260/280 nm (A260/A280) using a Nanodrop®ND-1000 spectrophotometer (LabTech, Holliston, MA, USA). The samples had an average RNA integrity number (RIN) value of 8.9, as determined using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). Small RNAs (18–30 nt) were gel-purified from 10 µg total RNA. Adaptors were ligated to eluted RNAs successively with RNA ligase. Libraries were gel-purified after each ligation step. Small RNAs ligated with adaptors were subjected to RT-PCR (15 cycles of amplification) to produce sequencing libraries. PCR products were purified on a non-denaturing acrylamide gel and sequenced for 50 cycles on Illumina HiSeq 2000 Sequencer (BGI, Shenzhen, China).
GEO Accession
GSM1363233