Template-switching during DNA synthesis by Thermus aquaticus DNA polymerase I

摘要: Recombinant DNA molecules are often generated during the polymerase chain reaction (PCR) when partially homologous templates available [e.g., see Paabo et al. (1990) J. Biol. Chem. 265, 4718-4721]. It has been suggested that these recombinant a consequence of truncated extension products annealing to on subsequent PCR cycles. However, we demonstrate here recombinants can be single round primer in absence heat denaturation, indicating template-switching produces some molecules. Two types template-switches were observed: (i) switches pre-existing and (ii) complementary nascent strand. Recombination is reduced several fold template strands physically separated by attachment streptavidin magnetic beads. This result supports hypothesis either or at least one two extending synthesis interaction between necessary for efficient template-switching.