GSM3684626: F508delCFBE41o- miRseq [Urban_2_T4_S22]; Homo sapiens; miRNA-Seq
Total RNA was isolated from cell lysate using the Quick-RNA Miniprep Kit (Zymo Research, R1054) according to the manufacturer's instructions, with additional on-column DNase treatment with the RNase-free DNase Set (Zymo) to remove contaminating genomic DNA. 100ng of total RNA was used with the QIAseqTM miRNA Library Kit (Qiagen, Maryland, USA) following manufacturer's instruction. 3' and 5' adapters were ligated to total input RNA. Reverse transcription, followed by PCR, was used to create cDNA constructs. PCR was performed with primers that anneal to the ends of the adapters. Quality was examined using Agilent Bioanalyzer Tapestation 2200 and Qubit 2.0 Fluorometer with miRNA library pre sequencing QC protocol mentioned in the QIAseqTM miRNA Library Kit. The cDNA libraries were pooled at a final concentration of 2.5pM. Cluster generation and 75 bp single read single-indexed sequencing was performed on Illumina NextSeq 500's.
GEO
GSM3684626
GEO Accession
GSM3684626