摘要:
The invention relates to the field of multiplex amplification. In particular, the invention relates to methods for assaying a sample for one or more nucleic acid targets in a single reaction based on the distinct melting temperatures or melting profiles of primers and/or probes. The invention also provides probes and kits for use in such methods.
摘要:
Disclosed herein are methods, compositions and kits for the quantification of a nucleic acid target present on a solid support. This entails quantitative real-time polymerase chain reaction wherein minor groove binders are excluded.
摘要:
Temperature control devices and methods are described. The described temperature control devices and methods comprise optical emission and detection assemblies and can be used in PCR and qPCR applications.
摘要:
This invention provides methods and systems for measuring the binding of analytes in solution to probes bound to surfaces in real-time. The method involves contacting a fluid volume having a plurality of different analytes with a solid substrate having a plurality of different probes. The probes are capable of specifically binding to the analytes. The method also involves measuring signals at multiple time points while the fluid volume is in contact with the substrate. The signals measured at multiple time points can be correlated with the amount of binding of the analytes with the probes. The method eliminates the need to wash the probes before measuring the binding characteristics.
摘要:
We describe a quantitative PCR (qPCR) instrument for combined qPCR and melt curve (dissociation and/or association curve) analysis. The instrument has at least one optical channel; a fluorescence excitation source; a fluorescence detector; an electronic analogue signal amplifier having an input coupled to an output of the fluorescence detector; and an analogue-to-digital converter (ADC) having analogue input coupled to an output of the analogue signal amplifier. The instrument further comprises a quantified automatic gain control (AGC) loop coupled between the signal output of the fluorescence detector and the analogue input of the ADC. The AGC loop is configured to apply a determined, numerical gain value to a fluorescence signal for the analogue input of the ADC. The instrument also includes a system to scale a digital output of the ADC responsive to the numerical gain value and to provide a digital fluorescence level signal from the scaled digital output.
摘要:
A method and device for digital multiplex PCR assays employ a microfluidic chip for performing real-time, continuous flow PCR within microchannels of the chip. A stream of sample material is introduced into each microchannel and alternating boluses of assay-specific reagents and buffer are introduced into the stream to form sequentially configured test boluses. A PCR procedure is performed on the test boluses followed by a thermal melt procedure. During the thermal melt procedure, fluorescent output is detected and fluorescence vs temperature data is collected and compared to expected normal correlations. The results, positive or negative, are converted to digital format, with positive results designated as “1” and negative results designated as “0”, or vice versa.
摘要:
The present invention features compositions and methods for quantifying detection of a target oligonucleotide in a sample in real time comprising one or more primer oligonucleotides comprising a 5′ nicking enzyme recognition site and a 3′-terminal region comprising a 2′-modified nucleotide. These methods are compatible with target oligonucleotides amplified using a nicking amplification reaction.
摘要:
A real-time quantitative PCR assay that utilizes a duplex, synthetic DNA standard to ensure optimal quality assurance and quality control. One embodiment of the invention facilitates amplification of mtDNA by focusing on a 105-base pair target sequence that is minimally homologous to non-human mtDNA. The present invention can also be used to identify the presence of PCR inhibitors and thus indicate the need for sample repurification.
摘要:
Described herein are microfluidic diagnostic methods and devices using electrokinetic modules for the isolation of targets (e.g., cells, bacteria, biomolecules) from biological samples, PCR amplification of DNA isolated from the targets, and real-time quantification of the amplified DNA using impedance sensing. Sample preparation, PCR amplification, and impedance sensing are thus performed using a single integrated platform.
摘要:
FRET-based analytes detection and related methods and systems are described where a pair of FRET labeled primers and/or oligonucleotides are used that are specific for target sequences located at a distance up to four time the Förster distance of the FRET chromophores presented on the FRET labeled primers and/or oligonucleotides one with respect to the other in one or more polynucleotide analyte; in particular the pair of FRET labeled primers and/or oligonucleotides is combined with a sample and subjected to one or more polynucleotide amplification reactions before measuring FRET signals from at least one FRET chromophore.