title: GSM3503239: CL214; Homo sapiens; miRNA-Seq
library_construction_protocol: Total RNA was extracted from peripheral blood mononuclear cells of CLL patients and 19+ B cells of healthy controls using the mirVana miRNA isolation kit (Thermofisher Scientific, MA, USA) . RNA quality/quantity and  miRNA quality/quantity was assessed using RNA 6000 Nano kit and small RNA kit respectively (Agilent technologogies, Santa Clara, USA) on Agilent 2100 bioanalyzer. For sequencing, libraries were generated using the IlluminaâTruSeqÔsmall RNA sample preparation kit (Illumina, San Diego, CA, USA). Briefly,  RNA adapters were ligated to 1 mg of total RNA. The ligated RNA was reverse transcribed to generate cDNA using RT primer, dNTP Mix, DTT, RNase Inhibitor and SuperScript II Reverse Transcriptase. The cDNA libraries were amplified using PCR to selectively enrich RNA fragments with adapter molecules. The resulting PCR products were purified using Ampure XP magnetic beads (Beckman Coulter) and run on a pre-cast agarose gel (Thermofisher Scientific, MA, USA). The gels were then extracted along the 160 bp RNA ladder band and the bottom of the 145 bp from the gel using the Mini Elute Qiagen Gel-Extraction Kit following the manufacturer's protocol (Qiagen, CA, USA). The quality and concentration of the libraries was assessed using the Qubit (Thermofisher Scientific, MA, USA) and DNA high sensitivity chip (Agilent Technologies, Santa Clara, USA). The libraries were then mixed at equimolar concentration.
attributes.tag: GEO Accession
attributes.value: GSM3503239