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1. Ansetzen des Primers an die DNA-Einzelstränge (spezifische Temperatur ca. 50-60°C)2. Synthese des komplementären Stranges in 5´-3´-Richtung durch die DNA-Polymerase.3. Wird zufällig ein Abbruchnukleotid eingebaut, so endet die Neusynthese.4. Da viele DNA-Einzelstränge im Ansatz sind und die Abbruchnukleotide zufällig eingebaut werden, ergibt sich im Ansatz ein Gemenge aus mehreren unterschiedlich langen DNA-Stücken, die aber alle mit dem gleichen Abbruchnukleotid enden.5. Gelelektrophoretische Auftrennung der 4 Ansätze. Hierbei wandern die kürzesten (also dem 5´Ende des neu synthetisierten Stranges am nächsten) DNA-Fragmente am weitesten.6. Um die Sequenz des DNA-Stranges zu ermitteln, muss man entsprechend noch vom Gel-Ende zu den Geltaschen in 3´- 5´-Richtung die Sequenz komplementär bilden.
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