GSM5737701: RNA-seq of population CD33+CD19+ of JIH-5 cells; Homo sapiens; RNA-Seq
RNA was extracted by ALLPrep Kit (Qiagen) Libraries were prepared from total RNA with the TruSeq Stranded Total RNA Library Prep Kit according to the manufacturer's instructions (Illumina, PN 20020599). For the low input:  Starting with 1-2 ng of total RNA,  cDNA was created using the Tecan/NuGen Ovation RNA Seq System V2 kit (7102-32).  Then 500 ng of the cDNA was sheared  to 300 bp with the Covaris LE220 (settings:  450 peak incident power (W), 15% Duty factor, 1000 cycles per burst, and treatment time of 100 seconds).  Libraries of the sheared cDNA was prepared using the Kapa Hyper library prep kit according to the manufacturer's instructions (KK8504). Total stranded RNA sequencing was performed by the Hartwell Center at St. Jude Children's Research Hospital. Libraries were prepared using the TruSeq Stranded Total RNA kit (Ilumina). 100 basepair, paired reads were generated by sequencing on the NovaSeq 6000 (Ilumina). For the low input:  Starting with 1-2 ng of total RNA,  cDNA was created using the Tecan/NuGen Ovation RNA Seq System V2 kit (7102-32).  Then 500 ng of the cDNA was sheared  to 300 bp with the Covaris LE220 (settings:  450 peak incident power (W), 15% Duty factor, 1000 cycles per burst, and treatment time of 100 seconds).  Libraries of the sheared cDNA was prepared using the Kapa Hyper library prep kit according to the manufacturer's instructions (KK8504).