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DNA-free A NEW Method to Remove DNA

A Isolation Method 1 single-reagent extraction method [e.g. TRIzol Reagent (Life Technologies), TRI Reagent (MRC), RNAzol (Tel-Test), RNA Stat-60 (Tel-Test), RNAwiz (Ambion)] 2 glass fiber filter-binding method [e.g. RNeasy (Qiagen), RNAqueous (Ambion)] 3 a multi-step guanidinium thiocyanate/acid phenol:chloroform extraction method (e.g. the Chomzynski and Sacchi procedure, Ambion's ToTALLY RNA Kit) 4 centrifugation through a CsCl cushion 5 two rounds of oligo d(T) selection [e.g. FastTrack RNA (Invitrogen), Poly(A)Pure Kit (Ambion)] 6 water control

Figure 1 shows that, regardless of the isolation method, gene specific product is synthesized in the absence of reverse transcriptase, indicating that none of these RNA isolation methods produce DNA-free RNA.


How Do You Know If Your RNA Is Contaminated with DNA?

How can you test for DNA contamination in RNA samples? The best way is to include a "minus-RT" control for each RNA sample in an RT-PCR experiment. If a PCR product is generated from an RNA sample that was not reverse transcribed (minus-RT control), then the product was amplified from contaminating DNA. Contaminating DNA may come from either the RNA preparation or from the RT-PCR reagents. A minus-template control for the PCR distinguishes between these possibilities.

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