Allelogic Biosciences Corporation

Biotium, Inc.

Xing Xin

Fei Mao

Wai-Yee Leung

Dimeric and trimeric nucleic acid dyes, and associated systems and methods are provided. Such a dye may form a hairpin-like structure that enables it to stain nucleic acids via a release-on-demand mechanism, for example. Such a dye may have low background fluorescence in the absence of nucleic acids and high fluorescence in the presence of nucleic acids, upon binding therewith, for example. A dye provided herein may be useful in a variety of applications, such as in DNA quantitation in real-time PCR, for example.

Dyes and labeled molecules

The present invention provides compositions, methods, and kits relating to the protection and deprotection of molecules comprising nucleophilic groups, such as the protection and deprotection of thermostable polymerases. Also provided are methods of performing nucleic acid amplification using polymerases protected according to the invention.

Compositions and methods for the protection of nucleophilic groups

Aspects of the invention relate to the use of novel oligonucleotides as temperature-sensitive inhibitors for thermostable DNA polymerases. Some inhibitors exhibit temperature-dependent and, in some cases, reversible inhibitory property by changing the conformation of at least a portion of the oligonucleotides from double-stranded to single stranded or in some cases vice versa in a temperature-dependent manner. Aspects also relate to the use of an the inhibitors in a hot-start PCR compositions, wherein the inhibitor may act to suppress the activity of the thermostable DNA polymerase below a desired activation temperature, Tact, and wherein the inhibitor is thermally inactivated above Tact, thus liberating the polymerase activity and initiating the DNA amplification process. Aspects further relate to a procedure for formulating the composition of a hot-start PCR reaction mixture. The hot-start PCR methods disclosed herein are generally faster, more flexible and lower in cost than existing methods.

Oligonucleotides as temperature-sensitive inhibitors for DNA polymerases

The present invention provides a dually labeled oligonucleotide probe, methods of preparing and using the same. The subject probes are particularly useful for high-sensitive nucleic acid detection via hybridization assays including but not limited to template-directed polymerization reactions.

Oligonucleotide probes and uses thereof

Dimeric and trimeric nucleic acid dyes, and associated systems and methods

An embodiment of the invention discloses new methods for designing labeled nucleic acid probes and primers by labeling oligonucleotides with a plurality of spectrally identical or similar dyes and optionally with one or more quencher dyes. Oligonucleotides labeled in accordance with some embodiments of the invention exhibit a detectable increase in signal, for example, fluorescent signal when the labeling dyes are separated from one another. Methods for separating the dye include cleaving the labeled oligonucleotides include using enzymes that have 5'-exonuclease activity. In one embodiment nucleic acid primers of the present invention may fluoresce upon hybridization to a target sequence and incorporation into the amplification product. Nucleic acid probes and primers of the present invention have wide applications ranging from general detection of a target nucleic acid sequence to clinical diagnostics. Major advantages of the oligonucleotides including nucleic acid probes and primers of many embodiments of the present invention are their synthetic simplicity, spectral versatility and superior fluorescent signal.

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Foster City, CA, United States of America

Xing Xin