GSM3932610: PSCSR-seq analysis for A549 cell line [A549_PSCSR]; Homo sapiens; miRNA-Seq
A549, Hela cultured cells, PBMCs, and B16F10 tumor cells were harvested and dissociated. Cells were dispensed into a ICELL8 nanowell chip for PSCSR-seq analysis. single cell small RNA analysis used PSCSR-seq protocol, briefly, small RNAs were ligated with 3' adapters using T4 RNA Ligase 2, truncated KQ (NEB), the remained 3' adapter were removed by 5' Deadenylase (NEB) and Lambda Exonuclease (Thermo Fisher). 5'adapters were ligated to the small RNA using T4 RNA Ligase 1(NEB). The adapters carrying 8 random nucleotides (UMI) for the small RNA counting. The ligated small RNAs were reverse-transcribed using Superscript III (Thermo Fisher), and two rounds of PCRs were performed to introduce the barcodes for labeling cells and multiplexing samples. PSCSR library was size selected with Pippin Prep, and the size distribution was assessed using an Agilent 2100 BioAnalyzer; Single cell RNA-seq followed Chromium Single Cell 3' Reagent Kits User Guide (v2 Chemistry) from the 10X Genomics company.