Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1993-04-19
1996-05-14
Jones, W. Gary
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
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:
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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).
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Backman Keith C.
Carrino John J.
Shimer George H.
Yocum Robert R.
Abbott Laboratories
Brainard Thomas D.
Jones W. Gary
Marschel Ardin H.
Yasger Paul D.
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