Amplification of nucleic acids
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United States of America Patent
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Jul 23, 2002
Grant Date -
N/A
app pub date -
May 15, 2000
filing date -
Aug 13, 1997
priority date (Note) -
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Abstract
The present invention relates to a method of amplifying a nucleic acid sequence present in a first strand of a double stranded nucleic acid molecule wherein said molecule incorporates an unmodified recognition site for a restriction enzyme capable of cutting the first strand at the 5' end of the sequence therein to be amplified and leaving the 3'-cut region of the second strand projecting beyond the cut site in the first strand. The method further comprises treating said molecule with said enzyme in the presence of a strand displacing polymerase and unmodified nucleotides for incorporation in an extending nucleic acid strand such that there is or becomes hybridised to said 3'-end region of the second strand a primer sequence complementary thereto whereby said primer sequence is extended in the 5' to 3' direction using the second strand as a template to regenerate the restriction endonuclease cut site and displace the sequence to be amplified.
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