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Primers with 5' flaps improve real-time PCR
Irina Afonina1, Irina Ankoudinova, Alan Mills
1Nanogen Inc., Bothell, WA 98021, USA. iafonina@nanogen.com
Biotechniques
|February 7, 2008
Abstract:
Primers that contain portions noncomplementary to the target region are usually used to add to the PCR product a utility sequence such as a restriction site or a universal probe binding site. We have demonstrated that primers with short 5'AT-rich overhangs increase real-time PCR fluorescent signal. The improvement is particularly significant for difficult to amplify templates, such as highly variable viral sequences or bisulfite-treated DNA.
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