PCR hot start using primers with the structure of molecular beacons (hairpin-like structure).
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.
Kaboev
O K
OK
St Petersburg Nuclear Physics Institute, Russian Academy of Science, Gatchina 188350, Russia and Tehran Pasteur Institute, Iran. kaboev@omrb.pnpi.spb.ru
Luchkina
L A
LA
Tret'iakov
A N
AN
Bahrmand
A R
AR
D020029:Base Pairing; D015374:Biosensing Techniques; D004396:Coloring Agents; D017931:DNA Primers / Q000737:chemistry* / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D005453:Fluorescence; D016158:Genes, p53 / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000235:genetics; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D019384:Nucleic Acid Synthesis Inhibitors; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D004396:Coloring Agents; D017931:DNA Primers; D019384:Nucleic Acid Synthesis Inhibitors; D004259:DNA-Directed DNA Polymerase