Methods of detecting polynucleotide kinase and its use as a...

Details Methods of detecting polynucleotide kinase and its use as a... Methods of detecting polynucleotide kinase and its use as a...

2006-10-31

2006-10-31

Horlick, Kenneth R. (Department: 1637)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

C435S091200, C435S091500, C435S091510, C536S025320, C536S025300

Reexamination Certificate

active

07129045

ABSTRACT:
Methods of detecting or measuring the activity of polynucleotide kinase are disclosed as well as methods of detecting an analyte in an assay using polynucleotide kinase as a label on a member of a specific binding pair. The methods rely on the phosphorylation of an oligonucleotide followed by ligation of the oligonucleotide 5′-phosphate onto another template-bound oligonucleotide. The presence of the ligated product signals the presence of polynucleotide kinase. In preferred embodiments, phosphorylation of an oligonucleotide enables the consecutive ligation of a set of oligonucleotides. The oligonucleotides so ligated can be detectably labeled with, for example, other enzymes to provide highly sensitive detection methods.

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C.E. Pritchard, E.M. Southern, Nucl. Acids Res., 25 3403-3407 (1997).

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