GSM6171830: NHBE cells, mock infected, rep 2; Homo sapiens; RNA-Seq
Cultures were dissected from transwell inserts, flash-frozen and stored in liquid Nitrogen. For harvesting, samples were added in a 1.5 ml Eppendorf tube containing 600 µl of Qiagen lysis buffer, and vortexed for 30 seconds at maximum speed. This dissociated cells from the membrane which was removed with tweezers. RNA was isolated using the RNeasy Mini Kit in combination with the RNase-free DNase Set as per manufacturer's instructions. RNA quantitation was performed using Nanodrop 2000 Spectrophotometer. The Illumina TruSeq Stranded mRNA HT kit (Illumina, 20020594) and SuperScript2 Reverse Transcriptase (Invitrogen, 18064014) as per manufacturer's instructions.