GSM8388850: B966, T cell lymphoma, biol rep 2; Mus musculus; RNA-Seq
Snap frozen tumor tissue was lyzed in homogenization buffer. Nuclei were pelleted by centrifugation (5 min, 4°C, 350 rcf). The supernatant was mixed with Trizol and chloroform, vortexed and centrifuged (15 min, 4°C, 21,000 rcf).  The aqueous layer was mixed with an equal volume of 100% ethanol and passed through a QIAgen RNeasy column. The QIAgen RNeasy protocol was followed and samples were eluted in 27 µL Elution Buffer. 3 µL of 10 x TurboTM DNase Buffer and 1 µL of TurboTM Dnase enzyme was added and the samples were incubated (30 min, 37°C). 70 µL of RNase-free water and 350 µL of QIAgen RLT Buffer were added. 250 µL of 100% ethanol was added and the samples were applied to the column and washed 2 x with RPE. Samples were eluted in RNase-free water. Quantseq, https://www.lexogen.com/quantseq-family/