title: GSM8211623: Neutrophils HD1; Homo sapiens; RNA-Seq
library_construction_protocol: Neutrophils were sorted directly into RLT-buffer and snap-frozen 2-7 ng of total RNA for each sample was prepared in a volume of 4 ul. Oligo dT primer hybridization was performed by the addition of Oligo dT mix (0.7ul H2O, 0.1 ul RNAse inhibitor (40U/ul), 0.1 ul dNTP mix (100mM) and 0.1 ul Oligo-dT30VN primer (100uM)). Reverse transcription was performed as described but the MgCl2 concentration was adjusted to 10 mM. Template switching and 10 (1-4 ng RNA input) or 11 (5-8ng RNA input) cycles pre-amplification of full length cDNAs with template switching oligo's was performed using ISPCR primer at a final concentration of 0.08 uM. The amplified full length cDNA was used for NGS library construction by Tagmentation for Illumina sequencing, using the Illumina Nextera XT DNA sample preparation kit (Illumina).