PCR hot start using primers with the structure of molecular beacons (hairpin-like structure) |
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| Authors: | Kaboev O K Luchkina L A Tret'iakov A N Bahrmand A R |
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| Affiliation: | O. K. Kaboev, L. A. Luchkina, A. N. Tret’iakov, and A. R. Bahrmand |
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| Abstract: | 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. |
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