title: GSM2065422: p53-KO-A1-1-C03_S1; Mus musculus; RNA-Seq
library_construction_protocol: Cell capture, lysis, reverse transcription and cDNA amplification was performed on the C1 IFC for mRNA-seq on Fluidigm C1 Single-Cell Auto Prep System following the manufacturer’s protocol. Medium sized C1 mRNA-Seq chips were used to capture each cell-cycle fraction. The C1 Auto-prep system captured the dissociated single cells across 96 wells and performed cell lysis, cDNA synthesis with reverse transcription and PCR reaction using the SMARTer Ultra Low Input RNA Kit. Cells captured across the 96 wells are manually inspected as a quality control measure to remove empty well, doublets or debris containing wells. 92 ERCC spike-ins of known concentration were added during the lysis step to assess and quantify technical variation. cDNA from several representative cells were checked by High Sensitivity DNA chips using Fragment Analyzer (Advanced Analytical). Libraries for each of the 96 captured cells were prepared using the Illumina Nextera XT DNA sample preparation kit with 96 dual barcoded indices. Single cell libraries were multiplexed and sequenced across 4 lanes of a NextSeq 500 platform (Illumina) using 75 single-end sequencing. On average, about 6-7 million reads would be generated from each single cell library.
attributes.tag: GEO Accession
attributes.value: GSM2065422