GSM8844357: DAOY cells with Hira loss-of-function treated with SAG_2; Homo sapiens; RNA-Seq
Daoy control and Hira KO cells were starved for 48 h with 500 nM SAG. Next RNA was isolated with the use of Universal RNA Purification Kit (EURx). RNA 3' mRNA-seq libraries were constructed using QuantSeq 3' mRNA-Seq Library Prep Kit reagents FWD for Illumina (Lexogen, cat. no. 015.96) and Lexogen i7 6 nt Index Set adapters (7001–7096; Lexogen, cat. no. 044.96). The manufacturer's protocol was followed, starting with 500 ng of supplied RNA and using 11 cycles of enrichment by amplification. The resulting 3' mRNA-seq libraries were subjected to fragment length control using an Agilent Bioanalyzer 2100 and High Sensitivity DNA kit reagents (Agilent, cat. no. 5067 − 4626). Library concentration was then determined by qPCR using the Kapa Library Quantification kit (Kapa Biosciences, cat. no. KK4824). The manufacturer's protocols were followed. Sequencing was carried out on an Illumina NovaSeq 6000 instrument using NovaSeq 6000 S1 Reagent Kit (200 cycles) reagents (Illumina, cat. no. 20012864), in pair-end read mode 2 × 100 cycles using the manufacturer's recommended standard procedure with 1% addition of Phix control library (Illumina, cat. no. FC-110-3001).