RNA-seq dRNA nanopore of Saccharomyces paradoxus, YPD
yeast cultures (2550 mL) were centrifuged at 1500 rpm for 3 min and washed with H2O, and cell pellets were immediately kept on ice. Each sample was then resuspended in 0.4 mL of AE buffer (50 mM sodium acetate at pH 5.3, 10 mM EDTA at pH 8.0). Sodium dodecyl sulfate was then added to a final concentration of 1%, and proteins and DNA were extracted by adding 0.6 mL of acidic phenol/chloroform (V/V), followed by incubation for 5 min at 65 degrees C. The aqueous phase was separated by centrifugation at 14,000 rpm for 2 min at 4 degrees C and washed with a volume of chloroform and separated by centrifugation at 14,000 rpm for 2 min at 4 degrees C. RNA was precipitated from the aqueous phase with ethanol. We subsequently performed poly(A)+ RNA purification using the NEBNext Poly(A) magnetic isolation module and concentration with the Monarch RNA cleanup kit
SUB16034013
S. paradoxus dRNA ONT