TURBO™ DNase removes 63-fold more plasmid DNA template from an in vitro transcription reaction than wild type DNase I.
In vitro transcription was accomplished using the MEGAscript® Kit. Following a 4 hr transcription reaction, RNA from triplicate reactions was treated with 1) no DNase I; 2) 2 U DNase I as per the standard protocol; 3) 2 U DNase I + DNase I buffer; or 4) 2 U TURBO™ DNase. The RNA in each sample was then purified with MEGAclear™ columns, eluted in nuclease-free water, and profiled on an RNA LabChip using Agilent 2100 bioanalyzer software. The concentration of nucleic acid was assayed using the NanoDrop® Spectrophotometer. The level of DNA contamination was quantified by real-time PCR using a TaqMan primer/probe set specific for the RNA transcript, and the respective cycle thresholds (Ct) were compared to a standard curve to determine the percent of residual, PCR-amplifiable DNA in each case.