Nested PCR

Sep 18, 2014

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Nested PCR. Nested PCR to metoda, w której stosuje si ę dwie pary starterów - zewn ę trzn e i wewn ę trz ne. Reakcja amplifikacji odbywa si ę w dw óch etapach.

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Nested PCR

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Nested PCR
NestedPCRto metoda, w której stosuje się dwie pary starterów - zewnętrzne i wewnętrzne. Reakcja amplifikacji odbywa się w dwóch etapach. Produkty powstałe w fazie pierwszej(preegzystujące produkty amplifikacji ), stają się następnie matrycą dla wewnętrznej pary primerów.
SCHEMAT METODY : * Wyizolowanie DNA * Pierwsza amplifikacja z zastosowaniem pierwszej pary primerów (położonych bardziej zewnętrznie) - ok. 40 cykli * Druga amplifikacja z zastosowaniem drugiej (położonej bardziej wewnętrznie – internal primers ) pary primerów – 15- 20 cykli

Zalety metody Nested PCR: * Mniejsza możliwość amplifikacji niepożądanych sekwencji * Podwyższona specyficzność * Podwyższona czułość

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