Next Generation Sequencing analyses negative control (NC) and TRPC5OS overexpressed MDA-MB-231 Transcriptomes

Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare the differential genes between negative control and TRPC5OS overexpressed MDA-MB-231. Method: RNA was extracted as aforementioned from cells following after TRPC5OS was overexpression.sed. Agarose gel electrophoresis was used for assessing the integrity testingof the extracted RNAs,, and quantification and further quality tests were performed using a NanoDrop ND-1000 (NanoDrop Technologies, Wilmington, DE, USA). Subsequently, an NEB Next ®Poly (A) mRNA Magnetic Isolation Module was used for mRNA enrichment by adding it into the total RNA extract. The library of the processed RNA product RNA was established using the KAPA Stranded RNA-Seq Library Prep Kkit (Illumina, Inc., San Diego, CA, USA). The established library was sequenced using the Illumina HiSeq 4000 platform. Conclusions: The results showed that total 55,256 genes were dysregulateddifferentially expressed, including 3,269 upregulated genes and 1,987 downregulated genes (fold change =1.5) . Overall design: two pairs of NC and TRPC5OS overexpressed MDA-MB-231