Becton Dickinson and Company Limited

Karen Eckert

Donald W Copertino

Matthew P Collis

Thomas L Fort

Tobin J Hellyer

Ray A McMillian

The present invention relates to methods for pretreating biological samples for extraction of nucleic acid therefrom. The present invention employs a combination of at least one protein denaturant with one or more of the following elements to form a reaction mixture for extraction of nucleic acid: (1) at least one aprotic solvent, (2) stepwise heating, and (3) sample dilution.

Pretreatment method for extraction of nucleic acid from biological samples and kits therefor

The present invention relates to methods for pretreating biological samples for extraction of nucleic acid therefrom The present invention employs a combination of at least one protein denaturant with one or more of the following elements to form a reaction mixture for extraction of nucleic acid: (1) at least one aprotic solvent, (2) stepwise heating, and (3) sample dilution.

For use in nucleic acid amplification reactions, the detector oligonucleotides of the invention comprise a target binding sequence which is at least partially the same as the target binding sequence of an amplification primer present in the target amplification reaction, so that the detector oligonucleotide and the amplification primer compete for hybridization to the same sequence in the target. Hybridization of the amplification primer to the target upstream from the detector oligonucleotide generates a nickable restriction endonuclease recognition site. When this site is nicked and strand displacement occurs from the nick, both the,&prime; end of the amplification primer and the detector oligonucleotide are displaced. The displaced detector oligonucleotide may then be detected as an indication of the presence of the target sequence, for example by unfolding of a fluorescently labeled secondary structure present in the detector oligonucleotide to reduce fluorescence quenching.

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Perry Hall, MD, United States of America

Karen Eckert