sRNA of CracrVA1 without Cas12a
CracrVA1 expressed from its native promoter was introduced into BW25113 . After culturing to the logarithmic growth phase, total RNA was extracted. A total of 50 g of RNA was treated with T4 polynucleotide kinase (NEB) according to the manufactures protocol. The RNA was purified using the phenol-chloroform method and then precipitated with an equal volume of isopropanol and 0.1 volume of 3M sodium acetate. The RNA molecules ranging from 30 to 300 nt were selected to construct a small RNA library with the NEXTFLEX Small RNA-Seq Kit