GSM3381019: Dif 10j A2 [JCZ_18]; Homo sapiens; ncRNA-Seq
Total RNA extraction was performed according to the Qiagen miRNeasy kit. RNA concentrations were determined using Qubit fluorometric quantification and purity by the RNA 6000 Bioanalyzer kit (Agilent). RNA-Seq libraries were constructed with the TruSeq stranded mRNA sample preparation (Low throughput protocol) kit and TruSeq Small RNA protocol from Illumina. The first step of the pipeline is transcripts purification by depleting abundant ribosomal RNAs with specific RiboZero kit. 3 μg of high quality total RNA was eluted in 10 μL RNase-free water for the construction of the libraries. All samples were supplied at the same concentration of 300 ng/μL and were DNase treated. The Illumina TruSeq Small RNA protocol is optimized to target small RNAs with 5'-monophosphate. This option is required for samples with unknown 5'-phosphorylation status. The final cDNA libraries were validated with a Fragment Analyzer (Advanced Analytical, Ankeny, IA) and quantified with a KAPA qPCR kit (Kapa Biosystems, Wilmington, MA). Having passed quality control, the libraries were sequenced on Illumina HiSeq 4000.
GEO
GSM3381019
GEO Accession
GSM3381019