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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.

PCR primers
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One-step, microplate or cuvet, colorimetric 530nm, detection limit 0.16mg/dL in serum or plasma. Procedure: 10 min....
MOUSE ANTI HUMAN NPM...
Edited by D. Rickwood, T.C. Ford and J.T. Steensgaard (1994) Compiles core data required by researchers carrying out any centrifugal separation, whether working with cells, organelles or macromolec...
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