摘要:
Methods for detecting a target polynucleotide sequences are provided that utilize a probe having a target-complementary segment and a detectable tag. By cleaving the detectable tab and associating the tag with a tag complement coupled to an electrode, an electrochemical signal can be detected that is related to the presence of the tag:tag complement complex.
摘要:
Methods and kits for detecting a target nucleic acid in a sample are described. In some embodiments, the sample to be analyzed includes a primer which hybridizes to at least a portion of the target nucleic acid, a probe having a first region which hybridizes to at least a portion of the target nucleic acid and a second region having a detectable label, a polymerase which extends the hybridized primer and an enzyme comprising nuclease activity that can cleave the hybridized hybridization probe to thereby release a labeled probe fragment. In some embodiments, the sample can then be contacted with a solid support comprising surface bound capture probes which can hybridize to the labeled probe fragment(s). These capture probes more readily bind to the probe fragment(s) than to the intact hybridization probe. The label can then be detected on the support surface. In this manner, improved discrimination between the probe fragments and the intact hybridization probes can be achieved.
摘要:
Methods and kits for detecting a target nucleic acid in a sample are described. In some embodiments, the sample to be analyzed includes a primer which hybridizes to at least a portion of the target nucleic acid, a probe having a first region which hybridizes to at least a portion of the target nucleic acid and a second region having a detectable label, a polymerase which extends the hybridized primer and an enzyme comprising nuclease activity that can cleave the hybridized hybridization probe to thereby release a labeled probe fragment. In some embodiments, the sample can then be contacted with a solid support comprising surface bound capture probes which can hybridize to the labeled probe fragment(s). These capture probes more readily bind to the probe fragment(s) than to the intact hybridization probe. The label can then be detected on the support surface. In this manner, improved discrimination between the probe fragments and the intact hybridization probes can be achieved.
摘要:
Microfluidic devices that incorporate a porous polymer electrode assemblies, including microfluidic device useful for detection of nucleic acids, as well as methods of using the microfluidic devices.
摘要:
Porous polymer electrode assemblies are useful in the detection or quantification of a variety of analytes. By preparing a porous monolith, and applying a conductive polymer to the monolith, a porous matrix is prepared that combines favorable conductive properties, by virtue of the presence of the conductive polymer, with the porous character of the underlying monolith. The resulting porous electrode can be used for qualitative or quantitative analysis, and the capture and/or release of selected charged materials, such as nucleic acids. The pores of the electrode matrix may also be filled with nonconductive material, yielding electrodes having a plurality of discrete conductive surfaces.
摘要:
The current teachings are directed to compositions, methods, and kits for selectively amplifying and for detecting target sequences. In some embodiments, a circularizable probe and/or a probe pair are disclosed for selectively amplifying target sequences. Methods for selectively amplifying target sequences are also disclosed, as are methods for detecting selectively amplified target sequences. Certain embodiments of the disclosed methods comprise a circularizable probe, a probe pair, comprising a first probe and a second probe, or both. In certain embodiments, a multiplicity of different circularizable probes, a multiplicity of different probe sets, or a multiplicity of different circularizable probes and a multiplicity of different probe sets are provided to selectively amplify or to detect a multiplicity of different target sequences, typically in a multiplex reaction. According to certain disclosed methods, surrogates of the target sequences are selectively amplified, including without limitation ligated probes, first amplification products, second amplification products, or combinations thereof. In some embodiments, selectively amplified target sequences or their surrogates are detected, directly or indirectly, indicating the presence of the corresponding target sequence. Kits to facilitate the performance of the disclosed methods are also provided.
摘要:
A diagnostic device is provided that includes a plurality of retainment regions, with the retainment regions that are separated by at least one dissolvable barrier. The retainment regions can be interconnected through at least one fluid processing passageway. A retainment region can include a container such as a retainment region, well, chamber, or other receptacle, or a retainment region such as a surface on which the material is retained. The retainment regions can include a reaction retainment region, one or more reagent retainment regions, each containing unreacted reagents, and a sample retainment region. A pressure-actuated valve can be positioned in each fluid processing passageway interconnecting the one or more reagent retainment regions with the respective intermediate retainment regions interposed between each of the one or more reagent retainment regions and the reaction retainment region. The dissolvable barrier can be a fluid flow modulator in the at least one fluid processing passageway.
摘要:
A diagnostic device is provided that includes a plurality of retainment regions interconnected through at least one fluid processing passageway or separated by at least one barrier. A fluid flow modulator can be provided in the fluid processing passageway if a fluid processing passageway is provided. The barrier and/or fluid flow modulator can comprise a polysaccharide, a derivative of a polysaccharide, or a combination thereof. For example, the barrier can comprise a chitosan material.
摘要:
A diagnostic device is provided that includes a plurality of retainment regions interconnected through at least one fluid processing passageway or separated by at least one barrier. A fluid flow modulator can be provided in the fluid processing passageway if a fluid processing passageway is provided. The barrier and/or fluid flow modulator can comprise a polysaccharide, a derivative of a polysaccharide, or a combination thereof. For example, the barrier can comprise a chitosan material.