( r by the competitors protocol. The trea...)The Fastest, Simplest, and Most Effective Way to Remove DNA Contamination

r by the competitors protocol. The treated RNA was subjected to real-time PCR to detect genomic DNA contamination that remained after DNase treatments. A GAPDH primer/probe set was used in triplicate reactions, and the extent of genomic DNA removal was calculated from the real-time PCR cycle threshold (Ct) values.

Elimination of DNA contamination by TURBO DNase, compared to no DNase treatment, shifted the real-time PCR results by >20 Ct values, representing >1.2 million fold reduction of DNA contamination. When compared to standard DNase I treatment, TURBO DNase decreased genomic DNA by an additional 22.6 fold (Figure 1). Indeed, one of the three TURBO DNA-free replicates was negative even after 40 cycles of PCR.

Figure 1. The TURBO DNA-free Kit Removes >20-fold more Genomic DNA than Competing Wild Type DNase I. RNA was isolated from mouse spleen using a procedure that results in significant contamination with genomic DNA. 8 g of the RNA was then treated with either 12 U of TURBO DNase or DNase I from Competitor P in a 100 l reaction. After DNase digestion, samples were processed by following the TURBO DNA-free or the competitors protocol. Triplicate samples of the treated RNA were subjected to real-time PCR using a GAPDH primer/probe set. Note that treatment with TURBO DNase shifted the average Ct threshold 4.5 cycles, equivalent to 22.6 fold reduction of genomic DNA contamination.

By comparison, the wild type DNase I removed 20 times less DNA; GAPDH was detected in all three replicates after 30 cycles. In addition, RNA LabChip analysis of the
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