GSM4227303: blood_gf_03; Mus musculus; RNA-Seq
Blood and tissue neutrophils were FACS sorted using CD45, CD11b, Ly6G and DAPI labelling, with typical purities > 95%. RNA was extracted ussing the RNAeasy plus micro kit from QUIAGEN cDNA amplification from neutrophil RNA and generation of index-tagged sequencing libraries were carried out using the Ovation Single Cell RNA-Seq System (NuGEN Technologies, San Carlos CA). Libraries were quantified using a Quant-iTTM dsDNA HS assay with the Q-bit fluorometer (Life Technologies, Carlsbad, California). Average library size and size distribution were determined using a High Sensitivity DNA assay in an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara CA). Libraries were normalized to 10nM using Tris-Cl 10mM, pH8.5 with 0.1% Tween 20. Libraries were applied to an Illumina flow cell for cluster generation (True Seq SR Cluster Kit V2 cBot) and 61 nt long, paired-end reads were generated on a Genome Analyzer IIx, using the TruSeq SBS Kit v5 (Illumina) and following the standard RNA sequencing protocol
GEO Accession
GSM4227303