GSM6622121: shKdm6a-GC1_RNA_A2; Mus musculus; RNA-Seq
Total RNA was extracted from pelleted cells by homogenizing in 1 ml of TRIzol reagent (Invitrogen, 15596026). After 5 mins of incubation, the lysate was mixed with 200µl of chloroform and centrifuged at 12,000 xg for 15 mins at 4°C. The aqueous phase was mixed with an equal volume of 100% ethanol and transferred to an RNeasy MinElute spin column (Qiagen, 74104) for purification according to the manufacturer's instructions. The quality of the eluted RNA was assessed by 28S/18S ribosomal ratios and RNA integrity number (RIN) using an Agilent Bioanalyzer. Libraries were constructed using the KAPA mRNA HyperPrep Kit (Roche # 08098123702) according to the manufacturer's instructions