Illumina HiSeq 2500 sequencing; Single cell RNA-sequencing of Spike-ins and Control RNA using 10x Genomics Chromium system
Single cell RNA-sequencing of Spike-ins and Control RNA using 10x Genomics Chromium system
An equal mix of Control Brain RNA (3µl; FirstChoice Human Brain Total RNA; #AM7962) and ERCC spikes (3µl 1:4 dilution; #4456653) was mixed to form ‘2x Control RNA+ERCC’ master mix. We further diluted ‘2x Control RNA+ERCC’ to ‘1x Control RNA+ERCC’ with PCR grade water. We made two single cell master mix preparation using 3µl of ‘2x Control RNA+ERCC’ and ‘1x Control RNA+ERCC’ respectively instead of single cell suspension (adjusted with 34.4 µl Nuclease-Free water). "The Single Cell Gel Bead kit (#120217), Single cell chip kit (#120219) and Single cell library kit (#120218) were used along with 10x GemCode Single Cell Instrument as per manufacturer specifications and manuals (Document # CG00011; Revision B)."