FAQ: Is it necessary to phosphatase treat my sgRNAs?

RNAs transcribed with a bacteriophage polymerase will possess 5’ triphosphates.  Phosphatase treatment will remove these triphosphates leaving a 5’ OH.  If treating with phosphatase we recommend treatment following DNaseI treatment.  The enzyme can not be inactivated by heat, therefore phenol:chloroform extraction is the preferred method for inactivating the enzyme.