title: RNA-Seq of Streptomyces venezuelae
design: totalRNA was treated by mRNA enrichment method or rRNA removal method. mRNA enrichment: magnetic beads with OligodT were used to enrich mRNA with polyA tail; rRNA removal: Hybridization of rRNA with DNA probe, selective digestion of DNA/RNA hybridization strand by RNaseH, digestion of DNA probe by DNaseI, purification of required RNA. The obtained RNA was segmented by interrupting buffer, and then reverse-transcribed by random N6 primers, and then synthesized cDNA double strand to form double-stranded DNA. The end of the synthetic double-stranded DNA is patched and phosphorylated at the 5 'end, forming A sticky end with a protruding "A" at the 3' end, and then connected to a bubbling splice with a protruding "T" at the 3' end. The linked products were amplified by PCR using specific primers. The PCR products are thermologically denatured into single strand, and then the single strand DNA is cycled with a bridge primer to obtain a single strand circular DNA library.