Hybridization mapping of nucleic acids

Details Hybridization mapping of nucleic acids Hybridization mapping of nucleic acids

1995-06-26

1997-01-07

Jones, W. Gary

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

935 78, 935 80, C12Q 168, C12N 1500

Patent

active

055915868

ABSTRACT:
A method is described which enables an investigator to locate where upon one single-stranded target nucleic acid molecule one or more separate single-stranded nucleic acid probe molecules hybridize. The method for determining where the probe hybridizes to the target is to allow the probe, present in excess relative to the target, and end-labeled (or labelable) target an opportunity to hybridize with one another after having been denatured together, if originally double-stranded, in a common reaction vessel and to then lightly digest the product of their hybridization with an agent that preferentially cleaves double- or single-stranded nucleic acids, depending upon the variation of hybridization mapping employed. The digestion should be such that, on average, each molecule which is cleaved is cleaved once. The cleaved reaction products are then fractionated by size to reveal the location of hybridization by reference to the pattern of signals produced by the label.

REFERENCES:
patent: 5217863 (1993-06-01), Cotton et al.
Galas and Schmitz (1987) Nucleic Acids Research 5:3157-3170.
Berk and Sharp (1977) Cell 12:721-732.
Kornberg and Baker, DNA Replication, 2nd edition (1992), p. 415.
Umthun et al. (1994) Nucleic Acids Research 22:4432-4440.
Hollingsworth et al. (1994) Nucleic Acids Research 22:1138-1146.
Wang et al. (1993) J. Biol. Chem. 268:10681-5.

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