Qiaseq targeted rna 96 index 1 kit

RNA was extracted from LSK cells using the micro RNA extraction kit (74034, QIAGEN) and from lymphoblast cell lines using the Mini RNA extraction kit (74134, QIAGEN) following the manufacturer’s instructions. RNA quality was assessed using an Agilent Bioanalyzer and RNA Nano Chip in the Biopolymers Facility of Harvard Medical School.
For gene expression analysis of LSK cells, cDNA was synthesized using the RT² PreAMP cDNA synthesis kit (33045, QIAGEN), target sequences were amplified using the RT² PreAMP Pathway primer Mix (PBM-029Z, 30241, QIAGEN) and qPCR Master Mix was RT² SYBR® Green (330533, QIAGEN). Multiplex real-time PCR was performed with the Mouse DNA damage signaling pathway PCR array (PAMM-029ZA, QIAGEN) following manufacturer’s instructions. PCR was performed in a QuantStudio 7 Flex real Time machine (Life Technologies). For the gene expression analysis in EUFA316 cells, a QIAseq Targeted RNA panel targeted was designed (CRHS-10510Z-219–12, 333022, QIAGEN) following manufacturer’s instructions. Index assignment was done using the QIAseq Targeted RNA 96-index I kit (333117, QIAGEN). Libraries were sequenced in a Illumina NextSeq 500 Mid Output flow cell and run in an Illumina NextSeq 500 sequencer in the Biopolymers Facility of Harvard Medical School.