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Nucleic Acids Research, 2000, Vol. 28, No. 21 e94
© 2000 Oxford University Press

PCR hot start using primers with the structure of molecular beacons (hairpin-like structure)

O. K. Kaboev*, L. A. Luchkina, A. N. Tret’iakov and A. R. Bahrmand1

St Petersburg Nuclear Physics Institute, Russian Academy of Science, Gatchina 188350, Russia and 1Tehran Pasteur Institute, Iran

A new technique of PCR hot start using oligonucleotide primers with a stem–loop structure is developed here. The molecular beacon oligonucleotide structure without any chromophore addition to the ends was used. The 3'-end sequence of the primers was complementary to the target and five or six nucleotides complementary to the 3'-end were added to the 5'-end. During preparation of the reaction mixture and initial heating, the oligonucleotide has a stem–loop structure and cannot serve as an effective primer for DNA polymerase. After heating to the annealing temperature it acquires a linear structure and primer extension can begin.

* To whom correspondence should be addressed. Tel: +7 812 71 46056; Fax: +7 812 71 32303; Email: kaboev@omrb.pnpi.spb.ru


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[Abstract] [Full Text] [PDF]


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