GSM7677099: MKN28_deltaTM; Homo sapiens; RNA-Seq
We performed RNAseq on total RNA from Luc, KIAA1324 WT-, 3NQ-, and ΔTM-expressing MKN28 cells. RNA quality was assessed by analysis of rRNA band integrity on an Agilent RNA 6000 Nano kit (Agilent Technologies, California, USA). Ahead of cDNA library construction, the 1ug of total RNA and magnetic beads with Oligo (dT) were used to enrich poly (A) mRNA from it. Then, the purified mRNAs were disrupted into short fragments, and the double-stranded cDNAs were immediately synthesized. The cDNAs were subjected to end-repair, poly (A) addition, and connected with sequencing adapters using the TruSeq RNA sample prep Kit (Illumina, California, USA). The suitable fragments that were automatically purified by BluePippin 2% agarose gel cassette (Sage Science, Massachusetts, USA) were selected as templates for PCR amplification. The final library sizes and qualities were evaluated electrophoretically with an Agilent High Sensitivity DNA kit (Agilent Technologies and the fragment was found to be between 350–450 bp.