System for multi color real time PCR

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 μm 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.