GSM8988871: 0.0 ug-Negative Control_4; Homo sapiens; RNA-Seq
Total RNA was extracted from cells using a mirVana™ RNA extraction kit (Life Technologies, Grand361 Island, NY).  RNA purity was assessed by using an RNA 6000 Nano LabChip™ Kit with a 2100 Bioanalyzer (Agilent Technologies,364 Palo Alto, CA). RNA sample concentrations were checked with a Qubit™ RNA BR Assay Kit. Poly (A) selection done using the Apollo 324™ System (IntegenX Inc., Pleasanton, CA). cDNA library prep was performed using the PrepX™ mRNA 48 366 Protocol on the Apollo 324™ System. PCR amplification took place across 14 cycles with 48 index primers, and the products were run through the Apollo 324™ PrepX™ PCR Cleanup 48 Protocol. The 2100 Bioanalyzer was used with a High Sensitivity DNA Chip to assess product quality and the resultant electropherogram was used to determine average size and concentration. A Qubit™ DNA HS Kit was then used to quantify the collected libraries. Equal amounts of each library were pooled into a total of 4 pools containing a maximum of 24 randomized libraries. The Qubit™ DNA HS Kit was again used to quantify the sequencing samples before they were denatured and diluted to a final concentration of 2.7 pM according to Illumina NextSeq 500 protocols. Samples were then sequenced using the Illumina NextSeq 500 (Illumina, San Diego, CA) with 75 single-read cycles