title: GSM3439553: SLX-11261.N702_S507.HGFMYBBXX.s_4.r_1; Homo sapiens; RNA-Seq
library_construction_protocol: The single cells were sorted in 96-well plates containing cell lysis buffer (2.3mL of 0.2% Triton X-100 with RNase inhibitor) and frozen to -80°C before library preparation. The method for single cell cDNA library making has been adapted from Smartseq2 for full-length RNAseq from single cells and Fluidigm (PMID: 24385147). Briefly, the reverse transcription was initiated with oligo-dT primers using SuperScript-II reverse transcriptase (Thermo Scientific) and supplemented with ERCC RNA Spike-In. The cDNA was then PCR amplified for 21 cycles using the KAPA HiFi Hotstart ReadyMix and IS primers (Kappa biosystems). After purification of the amplified cDNA, the indexed library was made using the Illumina Nextera XT DNA kit, 96 cells eventually pooled and library quality checked on an Agilent high sensitivity DNA chip before sequencing on one lane of an HiSeq-4000 (Illumina).
attributes.tag: GEO Accession
attributes.value: GSM3439553