GSM7987186: MT2 HBSS-4; Mus musculus; RNA-Seq
Approximately 3mm3 tissue was minced in 500uL buffer RP1 (Machery Nagel) + 1% bME + 0.5% antifoam Y-30 using M-tubes (Miltenyi Biotech), homogenized using a GentleMACS disruptor on the “RNA” protocol (Miltenyi Biotech). The lysate was passed through a 20Ga needle at least 5 times to produce the stock lysate. For RNA extraction, 200uL stock lysate was mixed with 500uL buffer RLT (Qiagen) homogenized further using a Qiashredder (Qiagen). The flow through was mixed 600uL 70% EtOH and loaded into a RNeasy spin column (Qiagen), then processed according to the manufacturer's instructions. PolyA selection was used to enrich for mRNA and RNA sequencing was performed by Azenta Life Science.