GSM4460010: HepG2_tRNA_siALKBH7-rep1; Homo sapiens; OTHER
Basically, as the first step of harvesting cells, the cells were washed once with DPBS buffer for each plate after aspirating the media. Then cellular total RNA was isolated with TRIzol reagent (Invitrogen) following manufacturer's protocol by isopropanol precipitation. The small RNA fraction (size less than 200 nt) was further extracted from the prepared cellular total RNA by mirVana™ miRNA Isolation Kit (AM1560, Invitrogen) protocols. Around 50 ng human cellular small RNA (size less than 200 nt) was fragmented into 40-50 nt using RNA Fragmentation Reagent (AM8740, Invitrogen) following the manufacturer's protocol and Oligo Clean & Concentrator (Zymo Research). These RNAs can be from diverse cellular treatments such as siALKBH7 vs. siControl or o/e Control vs. o/e ALKBH7. Then the library construction procedures will be conducted with T4 PNK end repair, RNA 3'-adapter ligation, HIV RT reverse transcription, cDNA 3'-adapter ligation and PCR amplification respectively, according to standard steps in section “DAOMM-seq for methylated site detection”. All libraries were sequenced on Illumina NovaSeq 6000 with single-end 100 bp read length.
GEO Accession
GSM4460010