Process for nucleic acid hybridization and amplification

Details Process for nucleic acid hybridization and amplification Process for nucleic acid hybridization and amplification

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01)

Patent

CA 2056983

The present invention is directed to a method of achiev- ing RecA protein facilitated amplification of a double- stranded DNA target sequence having first and second complementary strands, each strand with 5' and 3' ends. The method involves complexing a primer complementary to a 5' end region of the first strand and a primer complementa- ry to a 5' end region of the second strand with RecA pro- tein in the presence of ATP-y-S. The complexed primers are then reacted in a mixture also containing the target se- quence, all four dNTPs, RecA protein and DNA polymerase. The reaction is conducted below the temperature required for thermal dissociation of the two target strands and continued until a desired degree of amplification of the target sequence is achieved. The present invention further includes the cloning and identification of the coding sequences for the RecA protein of Thermus aquaticus.

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