Primers with 5' flaps improve real-time PCR.

  title={Primers with 5' flaps improve real-time PCR.},
  author={Irina Afonina and Irina Ankoudinova and Alan Mills and Sergey G. Lokhov and Phan Huynh and Walt Mahoney},
  volume={43 6},
          770, 772, 774
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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