title: GSM2194996: bulk RNA-seq M772LRC; Homo sapiens; RNA-Seq
library_construction_protocol: cDNA synthesis protocol: Cells were sorted into lysis buffer composed of 0.2 % Triton X-100 (Sigma) and 2 U/μl of RNAse Inhibitor (Life Technologies). ERCC spike-in controls (Life Technologies) were added to the cell lysis mix at 1:5,000 dilution. RNA was cleaned-up from the crude lysate with Agencourt RNAclean XP SPRI beads (Beckman-Coulter). cDNA was synthesized and pre-amplified from 5 μl of lysate according to the Smart-seq2 protocol (Picelli et al., 2013) 0.2-1 ng of pre-amplified cDNA was used as input for tagmentation by the Nextera XT Sample Preparation Kit (Illumina), where amplification was performed for 12-14 PCR cycles. Cells were isolated from the murine bone marrow, enriched by a MACS step and sorted for the population of interest. All steps were performed at 4°C.
attributes.tag: GEO Accession
attributes.value: GSM2194996