DNA sequencing by stepwise ligation and cleavage

Details DNA sequencing by stepwise ligation and cleavage DNA sequencing by stepwise ligation and cleavage

1996-06-21

1998-02-03

Jones, W. Gary

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 911, 435 912, 435 9152, 435 9153, 536 243, 935 77, 935 78, C12Q 168, C12P 1934, C07H 2104, C07H 2102

Patent

active

057143307

ABSTRACT:
The invention provides a method of nucleic acid sequence analysis based on repeated cycles of ligation to and cleavage of probes at the terminus of a target polynucleotide. At each such cycle one or more terminal nucleotides are identified and one or more nucleotides are removed from the end of the target polynucleotide, such that further cycles of ligation and cleavage can take place. At each cycle the target sequence is shortened by one or more nucleotides until the nucleotide sequence of the target polynucleotide is determined. The method obviates electrophoretic separation of similarly sized DNA fragments and eliminates the difficulties associated with the detection and analysis of spatially overlapping bands of DNA fragments in a gel, or like medium. The invention further obviates the need to generate DNA fragments from long single stranded templates with a DNA polymerase.

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