Studying the effect of single mismatches in primer and probe binding regions on amplification curves and quantification in real-time PCR.
Comparison of a broad range of characteristics of real-time PCR amplification curves yielded only slight alterations for low numbers of mismatches in the primer binding regions, resulting in a quantification error up to 63.12%. The effects were more pronounced for mismatches in the probe binding region and resulted in a quantification error up to 33%.
Süss
Beate
B
Institute for Milk Hygiene, Milk Technology and Food Science, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine Vienna, Veterinaerplatz 1, 1210 Vienna, Austria.
Flekna
Gabriele
G
Wagner
Martin
M
Hein
Ingeborg
I
D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D001431:Bacteriological Techniques; D020137:Base Pair Mismatch; D017931:DNA Primers / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D004269:DNA, Bacterial