RNA-seq for S. cerevisiae expresses MEP pathway from bacteria with two additional copies of IspC
Total RNA was extracted from yeast cells using the RNAprep Pure Plant Kit (Tiangen, China). RNA purity and concentration were quantified on a NanoDrop 2000 spectrophotometer (Thermo, USA), and RNA integrity was verified using an Agilent Bioanalyzer 2100 with RNA Nano 6000 reagents (RIN  8.5 for all samples). Libraries were constructed from 1 g total RNA per sample with the Hieff NGS Ultima Dual-mode mRNA Library Prep Kit (Yeasen, China). Polyadenylated mRNA was enriched using oligo(dT)-coupled magnetic beads, fragmented, and reverse-transcribed into double-stranded cDNA. After end repair, adenylation, and ligation of Illumina adapters, libraries were amplified with indexed primers and purified with AMPure XP beads (Beckman Coulter, USA). The sequencing was performed on the Illumina NovaSeq platform (151 bp paired-end reads).
SUB15284861
BQ7470022