GSM8071091: 6DT1 shH4 rep2; Mus musculus; RNA-Seq
For RNA-seq, cells were seeded at equal density in triplicate per condition. After 24 hours, media was aspirated and TriPure isolation reagent was added to the dish to collect cells before centrifugation to separate RNA and organic phases. After separation, the RNA phase was transferred to a new tube and isopropanol was added to the RNA to precipitate. The RNA isopropanol precipitation sample mixture was added to a Qiagen RNeasy Spin column for purification using the Qiagen RNeasy kit. Samples were analyzed by the Agilent 2200 TapeStation electrophoresis system. All samples used for RNA-seq had an RNA integrity number (RIN) score greater than 7 and were sent to the Illumina Sequencing Core at the Frederick National Labs for sequencing. Libraries were constructed using TruSeq Stranded Total RNA Library Prep, following the manufacturers protocol