Ethidium monoazide and propidium monoazide for elimination of unspecific DNA background in quantitative universal real-time PCR.
Unspecific background DNA in quantitative universal real-time PCR utilizing a hydrolysis probe was completely suppressed by the addition of EMA or PMA to the PCR mix via cross-linking of the dyes to DNA during 650 W visible light exposure. The proposed procedure had no effect on the sensitivity of the real-time PCR reaction.
Hein
Ingeborg
I
Institute of Milk Hygiene, Milk Technology and Food Science, Department of Veterinary Public Health and Food Science, University of Veterinary Medicine, Veterinaerplatz 1, A-1210 Vienna, Austria. ingeborg.hein@vu-wien.ac.at
Schneeweiss
Wilfried
W
Stanek
Christian
C
Wagner
Martin
M
D001386:Azides / Q000737:chemistry*; D004269:DNA, Bacterial / Q000032:analysis*; D007364:Intercalating Agents; D016133:Polymerase Chain Reaction / Q000379:methods*; D011419:Propidium / Q000737:chemistry*; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification*
D001386:Azides; D004269:DNA, Bacterial; D007364:Intercalating Agents; D011419:Propidium; C014535:8-azidoethidium