Method of preparing primers for polymerase chain reaction...

Details Method of preparing primers for polymerase chain reaction... Method of preparing primers for polymerase chain reaction...

C - Chemistry, Metallurgy

12

Q

150/8.5

C12Q 1/68 (2006.01) C12N 15/10 (2006.01)

Patent

CA 2034161

Abstract (a) A method of preparing primers for polymerase chain reaction, (b) a method of amplifying DNA in vitro by the polymerase chain reaction and (c) a method of amplifying a chromosomal gene. The method of preparing primers for polymerase chain reaction comprises treating a double-stranded DNA fragment with an exonuclease. The method of amplifying DNA in vitro by the polymerase chain reaction comprises using, as a primer, a DNA fragment obtained by treating a double-stranded DNA fragment with an exonuclease. The method of amplifying a chromosomal gene comprises carrying out the inverted polymerase chain reaction using, as a primer, a DNA fragment obtained by treating a double-stranded DNA fragment with an exonuclease capable of digesting one chain of the double-stranded DNA in a manner such that mononucleotides are released therefrom in the direction from the 5' terminus to the 3' terminus and, as a template, a circular DNA obtained by cleaving the chromosomal gene with a restriction enzyme and subjecting the resultant fragment to self-ligation.

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