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DNA-free A NEW Method to Remove DNA


Now it's easy to make your RNA free of genomic DNA contamination and ready for RT-PCR. DNA-free DNase Treatment & Removal Reagents contain RNase-free DNase, and an optimized DNase digestion buffer, to ensure safe, complete removal of contaminating DNA from any RNA sample. Also included is a unique DNase Removal Reagent which, after digestion, eliminates DNase in minutes no more messy phenol extractions or heat inactivation procedures which can cause RNA loss or degradation.

DNA, contaminating RNA preparations, can serve as a template in PCR to produce a false positive signal from RT-PCR. Although false positives are easily identified by looking at the outcome of a "minus-RT" control, eliminating the DNA is not trivial. Here we discuss the problem of genomic DNA contamination in RT-PCR, the best methods to detect and remove it, and an innovative way to remove DNase I after DNase I treatment.


All Isolation Methods Result in Contaminating DNA

There is no RNA isolation method that consistently produces RNA free from genomic DNA without the use of DNase. To illustrate this, RT-PCR was performed on mouse liver RNA isolated by several different methods:

Figure 1. DNA Contamination in RNA Isolated by 5 Different Methods. Total RNA was isolated from mouse liver by the methods indicated. RNA (0.5 g) underwent RT-PCR, or simply PCR (without reverse transcription), as indicated and aliquots of each reaction were electrophoresed on a 2% agarose gel and stained with ethidium bromide.

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