Rna-Seq of Healthy Control
Generation of mRNA libraries and their sequencing was carried out at the Lifecell International Pvt Ltds core facility in Bangalore. Total RNA was converted into double-stranded DNA libraries using the NEBNext First Strand Synthesis Enzyme Mix. This was followed by synthesis of second strand cDNA and purification of double stranded cDNA using Jetseq beads. cDNA library was subjected to End-repair and adapter ligation before PCR based selective enrichment of DNA fragments in a 12-cycle reaction, followed by purification using Jetseq beads. Final purified mRNA libraries were assessed for fragment size distribution using Tapestation 4200 system (Agilent). High-throughput sequencing was carried out with a NovaSeq 6000 Sequencing System (150bp paired-end reads).