System for multi color real time PCR
First Claim
Patent Images
1. A real time PCR instrument comprising:
- an excitation unit comprising;
a plurality of reaction vessels for containing a reaction mixture;
exactly one monochromatic light source, wherein the monochromatic light source is an LED emitting at 470 nm; and
a lightpipe arranged to receive light from the reaction vessels and distributing said light into a six-leg optical fiber bundles of 50 μ
m single glass fibers;
a detection unit comprising six separate fluorescent detector entities, each of said detector entities having a central detection wavelengths at 530 nm, 555 nm, 610 nm, 640 nm, 670 nm, and 710 nm +/−
5 nm respectively, wherein the excitation unit and the detection unit are mechanically decoupled and connected by the six-leg optical fiber bundles where each bundle transmits light into each of the six detector entities; and
means for heating and cooling;
wherein the detector entities simultaneously detect maximum fluorescence emission of at least four differently labeled FRET hybridization probe pairs, simultaneously detect maximum fluorescence emission of at least two differently labeled TaqMan hybridization probes, and detect maximum fluorescence emission of SybrGreenI.
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Abstract
The invention is directed to a system for performing multi-color real time PCT, comprising a flexible real time PCT 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.
31 Citations
4 Claims
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1. A real time PCR instrument comprising:
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an excitation unit comprising; a plurality of reaction vessels for containing a reaction mixture; exactly one monochromatic light source, wherein the monochromatic light source is an LED emitting at 470 nm; and a lightpipe arranged to receive light from the reaction vessels and distributing said light into a six-leg optical fiber bundles of 50 μ
m single glass fibers;a detection unit comprising six separate fluorescent detector entities, each of said detector entities having a central detection wavelengths at 530 nm, 555 nm, 610 nm, 640 nm, 670 nm, and 710 nm +/−
5 nm respectively, wherein the excitation unit and the detection unit are mechanically decoupled and connected by the six-leg optical fiber bundles where each bundle transmits light into each of the six detector entities; andmeans for heating and cooling; wherein the detector entities simultaneously detect maximum fluorescence emission of at least four differently labeled FRET hybridization probe pairs, simultaneously detect maximum fluorescence emission of at least two differently labeled TaqMan hybridization probes, and detect maximum fluorescence emission of SybrGreenI. - View Dependent Claims (2, 3, 4)
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Current AssigneeRoche Molecular Systems (Roche Holding AG)
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Original AssigneeRoche Molecular Systems (Roche Holding AG)
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InventorsSagner, Gregor, Bechler, Ingrid, Bolte, Jochen, Heindl, Dieter, Josel, Hans-Peter, Gutekunst, Martin, Sebl, Rudolf, Mueller, Christoph
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Primary Examiner(s)Strzelecka, Teresa E
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Application NumberUS10/549,648Publication NumberTime in Patent Office2,910 DaysField of SearchNoneUS Class Current422/50CPC Class CodesC12Q 1/6851 Quantitative amplificationC12Q 2537/143 Multiplexing, i.e. use of m...C12Q 2561/113 Real time assayC12Q 2565/101 Interaction between at leas...G01N 2021/6421 Measuring at two or more wa...
