GSM4610556: mIDH1_rep2 [72865_GGCTAC_S10_L001]; Mus musculus; RNA-Seq
Tumor was dissected and processed to obtain a single cell suspension. Immune cells were enriched by percoll purification. Myeloid cells were sorted by flow cytometry. Total RNA was isolated from the sorted tumor-infiltrating CD11b+ Gr-1+ cells using RNeasy Plus Mini Kit RNA quality was assessed using a TapeStation (Agilent Technologies) following the manufacturer's recommended protocol. Samples with RINs (RNA Integrity Numbers) of 8 or greater were prepared using the Illumina TruSeq mRNA Sample Prep Kit v2 (Illumina, RS-122-2001, RS-122-2002). An amount of 0.1-3 μg of total RNA was converted to mRNA using polyA purification. The mRNA was then fragmented and copied into first-strand cDNA using reverse transcriptase and random primers. The 3' ends of the cDNA were then adenylated and ligated to adapters. Each ligated adapter had a unique 6 nucleotide barcode to identify each sample, enabling multiple samples to be sequenced from each lane of a HiSeq flow cell (Illumina). The products were purified and enriched by PCR to create the final cDNA library. Libraries were checked for quality and quantity by TapeStation (Agilent Technologies) and qPCR using Kapa's library quantification kit for Illumina Sequencing platforms (Kapa Biosystems, KK4835) using the manufacturer's recommended protocols.
GEO Accession
GSM4610556