GSM5740351: miRNA-seq Neu WT rep 1; Homo sapiens; miRNA-Seq
To calculate relative and absolute differences in the miRNA population in NEUWT and NEURTT, small RNAs were extracted from two replicates of both lines using the same number of cells followed by the addition of a set of spike-in RNAs. Small RNAs were extracted from 500,000 neurons of each line using the SPLIT RNA extraction Kit (Lexogen) according to the manufacturer's instructions. A set of 52 RNA spike-ins (QIAseq miRNA Library QC Spike-Ins – Qiagen) that spanned a wide range of concentrations were added to the recovered RNAs according to the manufacturer's instructions. Sequencing libraries were made using the Small RNA library preparation kit NEBNext (NEB) according to the manufacturer's instructions. Sequencing was performed on the Illumina HiSeq 2500 using the Rapid Run mode.