RNAseq of S. albidoflavus/scl sclM4 replicate 2
Streptomyces were grown on 33 mL of supplemented minimal media solid(SMMS) for 120 hours. The cell biomass was scraped off and collected into graduated conicaltubes with an appropriate amount of bashing beads, appropriate amount of 1X DNA/RNAProtection Reagent, Prot K Reaction Buffer and Prot K from Monarch Total RNA Miniprep Kit(New England Biolabs). After incubating at 55 C for 10 minutes, the cell biomass washomogenized with VelociRuptor V2 Microtube Homogeniser at 6.0 m/s for 40 seconds. Celllysis, RNA binding and eluting were performed followed the manufactures protocol.After the extraction, 200 ng of extracted RNA was run on 1% agarose gel to test theintegrity. PCR was performed to detect gDNA contamination. Turbo DNase (Fisher Scientific)was used to eliminate any potential gDNA contamination.