Methods for labeling DNA ends with halogenated nucleotides and detecting same with antibodies
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United States of America Patent
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Jun 15, 1999
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N/A
app pub date -
Oct 22, 1996
filing date -
Aug 30, 1995
priority date (Note) -
Expired
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Abstract
The invention pertains to the field of DNA detection for basic research, medical diagnostic testing, and forensic testing. Methods are provided for end labeling of DNA strands without a denaturation step so that cellular morphology can be better preserved. The DNA strands are first incubated with a halogenated deoxynucleotide triphosphate, such as brominated deoxyuridine triphosphate (BrdUTP), and an enzyme which can catalyze the addition of the halogenated deoxynucleotide to the 3' OH ends of the DNA strand, such as terminal deoxynucleotidyl transferase (TdT). The resulting modified DNA strands are then incubated with a labeled antibody, such as a fluoresceinated monoclonal antibody, that binds specifically to the halogenated deoxynucleotide. The label is then detected, e.g., by flow cytometry. The methods have utility in detecting apotosis, DNA synthesis and/or repair, and as general methods for end labeling DNA.
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