protein gfp and a polymerase was created via recombinant techniques well known in the art quartz chips 14 mm in diameter 0 3 mm thick were spin coated with a layer of planar dextran modified with immobilised streptavidin xantec germany an upright microscope 135tv zeiss equipped with total internal reflection tir illumination served as a platform for the experiment a laser beam of peak 488 nm melles griot with a power of 100 mw was circularly polarised by quarter wave plates to undergo tir in a custom made prism the prism was coupled optically to the chip via index matching gel a flow cell with an o ring seal and transparent optical axis was then assembled over the prism and quartz chip an objective collected the fluorescence signal through the transparent top surface of the flow cell images traveled via a band pass filter into a back thinned charge coupled device andor technologies two oligonucleotides were synthesized using standard phosphoamidite chemistry the oligonucleoitde defined as seq id no 1 was used as the target polynucleotide and the oligonucleoitde defined as seq id no 2 was biotinated and used as a primer the biotinated primers seq id no 2 where deposited onto the streptavidin coated chip by injection into the flow cell 10 pm of seq id no 2 in tris buffer containing 100 mm mgcl 2 and allowed to incubate for 10 minutes the flow buffer was flushed into the flow cell at a rate of 500 u03bcl min the complementary sequence seq id no 1 was then injected into the flow cell at a concentration of 1 u03bcm at a flow rate of 5 u03bcl min during this.