Methods for nucleic acid detection, sequencing, and cloning usin

Details Methods for nucleic acid detection, sequencing, and cloning usin Methods for nucleic acid detection, sequencing, and cloning usin

1995-06-07

1998-04-28

Jones, W. Gary

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 5, 435 912, 435 911, 435174, 424 941, 536 266, 536 231, 536 243, 536 2433, C12Q 168, C12P 1934, C07H 2102, C07H 2104

Patent

active

057443068

ABSTRACT:
The present invention provides a method of detecting the presence of a nucleotide sequence within a double-stranded DNA in a sample comprising: a. digesting the double-stranded DNA with an exonuclease which converts at least a portion of the double-stranded DNA to single-stranded DNA; b. hybridizing the single-stranded DNA with i) a first nucleic acid probe adapted with a moiety which can be captured to a solid support, and ii) a second nucleic acid probe labeled with a detectable moiety which can hybridize with the single-stranded DNA adjacent the hybridized first nucleic acid probe; c. ligating the hybridized first and second nucleic acid probes; d. capturing the moiety on the first nucleic acid probe hybridized to the DNA to the solid support; e. denaturing the ligated first and second nucleic acid probes from the hybridized single-stranded DNA; f. removing uncaptured labeled probe; and, g. detecting the presence of captured detectable moiety, the presence of captured detectable moiety indicating the presence of the nucleotide sequence within the double-stranded DNA in the sample.

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