Transcriptome Profiling of UCL93 Kidney Epithelial Cells and CRISPR-Cas9 Knockouts for PKD1 and BICC1 Using Total RNA-Seq
Transcriptome Profiling of UCL93 Kidney Epithelial Cells and CRISPR-Cas9 Knockouts for PKD1 and BICC1 Using Total RNA-Seq
UCL93 and KO cells were plated at 5x10^5 cells/mL in 10 cm2 plates and incubated at 33°C under 70% confluency was reached. Total RNA was extracted from the cells (n=3/ cell line).  Total RNA was extracted from the cells (n=3/ cell line) using a Trizol reagent and stored at -80°C. The RNA concentration and quality were determined using a Thermo Scientific™ NanoDrop™ spectrophotometer and Agilent 2100. After rRNA removal at Novogene, the mRNA is fragmented randomly by adding fragmentation buffer, then the cDNA is synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. Second, after a series of terminal repair, A ligation and sequencing adaptor ligation, the double-stranded cDNA library is completed through size selection and PCR enrichment.