The World's Best DNase
|Improved TURBO DNA-free™ |
The Best Way to Remove Contaminating DNA
Enhanced DNase Activity
Figure 1. DNA Removal Improved >600 fold with New TURBO DNA-free™. Mouse spleen total RNA samples, highly contaminated with DNA (30% RNA and 70% DNA; 23 µg), were treated with 4 U of TURBO DNase in a 60 µl reaction for 20 min at 37°C, or were left untreated. DNase digestion was halted by adding 6 µl (1/10 volume) of DNase Inactivation Reagent. Each treated sample (2 µl) was amplified in a 25 µl RT-PCR using a TaqMan® primer:probe set for mouse GAPDH. RT-PCR analysis of the DNase treated samples unmasked the RNA-only signal, which appeared at 15.3 Ct.
Additional studies have corroborated the superior efficiency of the improved kit in eliminating DNA from different sources of RNA using a variety of primer:probe sets in RT-PCR (data not shown). As a result, the TURBO DNA-free Kit, already the best choice for eradicating DNA from RNA preparations, is now even more effective in digesting DNA away from the most contaminated samples.
Destroy DNA and Preserve RNA Quality
Maximum RT-PCR Sensitivity
At high cycle numbers, RT-PCR is significantly less robust than it is in earlier cycles; suboptimal salt or pH conditions, or contaminants that have little impact on accurate detection at 15-25 cycles can profoundly distort detection and quantitation at >30 cycles. For this reason, the improved TURBO DNA-free Kit was carefully tested at the lower threshold for target detection to ensure maximum RT-PCR responsiveness. As shown in Figure 2, transcript levels from as little as 1 pg of total RNA could be quantitated in one-step qRT-PCR within a single Ct of the untreated control reaction. Thus, TURBO DNA-free offers researchers the confidence that rapid, simple, and highly effective DNA and DNase removal does not compromise sensitivity.
Figure 2. Treatment of RNA with TURBO DNA-free™ Maintains Target Sensitivity in Real-time PCR. (A) Purified HeLa total RNA (100 pg and 1 pg) was treated with the improved TURBO DNA-free according to the standard protocol. 5 µl of the treated sample was reverse transcribed with Ambion’s MessageSensor™ RT Kit and then amplified with a human b-actin TaqMan® primer/probe set in one-step RT-PCR. (B) The same analysis described above was performed, except that PCR amplification was monitored with a human CDC-2 TaqMan primer/probe set.