System For Multi Color Real Time PCR
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Abstract
The invention is directed to a system for performing multi-color real time PCR, comprising a flexible real time PCR instrument and a specific composition or reaction mixture for performing multiplex PCR: In particular, the present invention is directed to a composition or reaction mixture which comprises at least 3, preferably 4-5 and most preferably exactly 4 pairs of FRET hybridization probes. Each pair of said hybridization probes consists of a FRET donor probe carrying a FRET donor moiety and a FRET acceptor probe carrying a FRET acceptor moiety having an emission maximum between 550 and 710 nm.
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Citations
27 Claims
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1-17. -17. (canceled)
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18. A real time PCR instrument comprising:
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One optical part for detecting fluorescence emission from a plurality of reaction vessels, the one optical part comprising; an excitation unit comprising; at least 1 light source capable of emitting light toward the reaction vessels containing fluorescent compounds, a lightpipe being arranged for receiving light from the reaction vessel and capable of distributing homogeneously said light into optical fiber bundles, wherein the light does not pass through a wavelength excluding device prior to being distributed into said optical fiber bundles, a detection unit comprising at least 5 separate fluorescent detector entities, each of said detector entities having a central detection wavelength, said wavelengths being distinct from each other by at least 25 nm, a plurality of at least 5 optical fiber bundles, each of said bundle being arranged for-receiving homogeneously distributed light from the lightpipe, and transmitting said light to said fluorescent detector entities, and means for heating and cooling, wherein the excitation and detection units are located in separate housings inside the one optical part enabling highly precise positioning of the excitation unit towards each of the reaction vessels without moving the detection unit. - View Dependent Claims (19, 20, 21, 22)
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23. A real time PCR instrument comprising:
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One optical part for detecting fluorescence emission from a plurality of reaction vessels, the one optical part consisting of; an excitation unit comprising; at least 1 light source capable of emitting light toward the reaction vessels containing fluorescent compounds, a lightpipe being arranged for receiving light from the reaction vessel and capable of distributing homogeneously said light into optical fiber bundles, wherein the light does not pass through a wavelength excluding device prior to being distributed into said optical fiber bundles, a detection unit comprising at least 5 separate fluorescent detector entities, each of said detector entities having a central detection wavelength, said wavelengths being distinct from each other by at least 25 nm, a plurality of at least 5 optical fiber bundles, each of said bundle being arranged for-receiving homogeneously distributed light from the lightpipe, and transmitting said light to said fluorescent detector entities, and means for heating and cooling, wherein the excitation and detection units are located in separate housings inside the one optical part enabling highly precise positioning of the excitation unit towards each of the reaction vessels without moving the detection unit. - View Dependent Claims (24, 25, 26, 27)
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Specification