RNAseq of streptomyces: whole organism
Total RNA was isolated, and the RNA_seq strand_specific libraries were constructed using the VAHTS Total RNA_seq (H/M/R) Library Prep Kit (Vazyme, China) according to the manufacturers instructions. RNA was purified by magnetic beads after removal of rRNA. Then RNA was fragmented into small pieces using divalent cations and the cleaved RNA fragments were copied into first strand cDNA using reverse transcriptase and random primers. Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. These cDNA fragments then go through the end repair process, addition of a single A base, and ligation of the adapters. The products were purified and enriched by PCR to create the final cDNA library.