GSM9267371: RNA-Seq, 95D cells, control; Homo sapiens; RNA-Seq
The full-length DAMER lncRNA sequence was transcribed in vitro using the T7 High Yield RNA Synthesis Kit. The transcribed RNA was 3'-end biotinylated using the Pierce RNA 3' End Biotinylation Kit to create the bait probe. Cell lysates were prepared from 95D cells using Pierce IP Lysis Buffer. 50 pmol of biotinylated RNA probe was added to streptavidin magnetic beads and incubated at room temperature for 30 minutes. The probe-bound beads were then incubated with 100 µl of a Master Mix containing the cell lysate for 1 hour at 4°C. RNA bound to the DAMER probe was eluted and purified using TRIzol reagent. Libraries were constructed from the extracted total RNA using standard protocols, including rRNA depletion, fragmentation, cDNA synthesis, and adapter ligation. The Qualified libraries were pooled and sequenced on Illumina platforms with PE150 strategy in Novogene Bioinformatics Technology Co., Ltd (Beijing, China), according to effective library concentration and data amount required.