Ligase chain reaction with endonuclease IV correction and contam

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical

Patent

Rate now

  [ 0.00 ] – not rated yet Voters 0   Comments 0

Details

435 6, 435 911, 436501, 536 221, 536 231, 536 241, 536 243, 536 2431, 536 2432, 536 2433, 935 77, 935 78, C12P 1934, C07H 2104

Patent

active

055166636

ABSTRACT:
The present invention involves methods of improving the Ligase Chain Reaction (LCR.TM.) amplification schemes by modifying at least one probe end so that the probability of the probe contributing to spurious ligation and signal development is greatly reduced. Only after specific hybridization of the modified probe with true target are the modified ends "corrected" by endonuclease IV in a target dependent fashion to allow participation of the probe in the enzymatic ligation reaction. Specific modifications include 3' phosphate blocking groups and nucleic acid overhangs containing an abasic site at the point of ligation. Further embodiments include probes modified to contain ribonucleotide moieties which, after amplification, can be cleaved by RNase to destroy the amplification products and reduce the risk of contamination.

REFERENCES:
patent: 4851331 (1989-07-01), Vary et al.
Levin, et al., "Metalloenzymes in DNA Repair", J. Biol. Chem. 266(34):22893-22898 (1991).
Johnson and Demple, "Yeast DNA Diesterase for 3' Fragments of Deoxyribose: Purification and Physical Properties of a Repair Enzyme for Oxidative DNA Damage", J. Biol. Chem. 263(34):18009-18016 (1988).
Siwek, et al., "The Relative Importance of Escherichia coli Exonuclease III and Endonuclease IV for the Hydrolysis of 3'-Phosphoglycolate Ends in Polydeoxynucleotides", Nucl. Acids Res. 16:5031-5038 (1988).
Doetsch and Cunningham, "The Enzymology of Apurinic/Apyrimidinic Endonucleases", Mutation Research, 236:173 -201 (1990).
Brutlag et al., J. Biol. Chem., 247, (1), (Jan., 1972), pp. 241-248.
Bailly et al., Biochem. J., 259, (1989), pp. 761-768.

LandOfFree

Say what you really think

Search LandOfFree.com for the USA inventors and patents. Rate them and share your experience with other people.

Rating

Ligase chain reaction with endonuclease IV correction and contam does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Ligase chain reaction with endonuclease IV correction and contam, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Ligase chain reaction with endonuclease IV correction and contam will most certainly appreciate the feedback.

Rate now

     

Profile ID: LFUS-PAI-O-1895057

  Search
All data on this website is collected from public sources. Our data reflects the most accurate information available at the time of publication.