GSM3142494: TN_24_5; Mus musculus; RNA-Seq
Liver was removed, flash frozen in liquid nitrogen and RNA was harvested using Bioline RNA Extraction kit. We adapted Cel-Seq2 protocol to sequence RNA transcripts in livers of mice treated with LPS. Cel-Seq2 was performed using reverse-transcription primers that contained anchored polyT, a 6 base pair unique barcode, a 6-base pair UMI (unique molecular identifier), the 5' Illumina adaptor as used in the Illumina small RNA kit, and a T7 promoter. Instead of single cells, total RNA isolated from liver was used for linear mRNA amplification.A total of 10 ng of RNA was used for library preparation, and the aRNA was converted to cDNA with 3' RNA adaptors sequences (RPI) found in the TruSeq Small RNA Library Prep Pooling Guide. A total of 11 cycles of PCR were performed with an elongation time of 30 seconds. Following cDNA cleanup and dilution, samples were multiplexed and sequenced on Illumina Nextseq 500 using Mid Output v2 Kit. Cel-Seq2 (modified for tissue)
GEO Accession
GSM3142494