GSM4680432: VC_No UV Round 1; Homo sapiens; RNA-Seq
Following treatment protocol, cell media was aspirated and each well was washed with PBS. 350 µL of lysis buffer (included in Gen Elute™️ Total RNA Purification Kit) was added. Wells were coated in lysis buffer for 5 minutes and transfered to seperate microcetrifuge tubes. 200 µL absolute ethanol was added and tubes were vortexed for 10 seconds. Columns were assembled with elution tubes and the lysates were applied directly onto seperately assembled columns. Tubes were centrifuged at 3,500 g for 1 minute. If lysate hasn't passed through the column. Centrifuge an additional minute at 14,000 g. Flowthrough was discarded. 400 µL of wash solution A was applied to each column and centrifuged for 1 minute at 3,500 g (or increased to 14,000 g as necessary). Flowthrough was discarded and the column was washed twice more with a 2 minute centrifuge time for the last wash. The columns were placed into fresh elution tubes and 50 µL of elution solution was added to each. The columns were centrifuged at 200 g for 2 minutes, followed by 14,000 g for 1 minute. The elution procedure was repeated to maximize yield. Samples were stored at -80 C until shipping at -20 C. mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. The mRNA was fragmented by addition of fragmentation buffer. NEB library and strand specific libraries were then chosen. (Novogene Corporation - protocol associated with NEB library and strand specific library) Libraries were constructed using standard Illumina protocol.
GEO Accession
GSM4680432