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6.

a11


    6.1.

    - 300 nM ;
    ( TaqMan) - 250 nM;
    MgCl2 - 5 nM (SYBR Green), 3.5 nM (TaqMan);
    - AmpliTaqGold, Applied Biosystems.


    6.2. SYBR Green

    AmpliTaqGold, Applied Biosystems
    18 min94OC
    230 sec94OC
    320 sec60-65OC
    41 min72OC
    5Plate read
    6Goto 2 for 39 times
    7Melting curve from 60OC to 94OC read every 0.2OC hold for 1 sec before plate read
    8Forever10OC


    1. , , . , 15-20 , .
    2. (Plate read) 72OC, ( . ).
    3. SYBR Green, , C(T) 35-38, 40 .
    4. AmpliTaqGold 60OC, . 3 4 1 min & 60-65OC, .
    5. (melting curve) SYBR Green PCR. , PCR- .


    6.3. TaqMan

    AmpliTaqGold, Applied Biosystems
    110 min94OC
    215 sec94OC
    41 min60OC
    5Plate read
    6Goto 2 for 39-45 times
    8Forever10OC


    1. Applied Biosystems -N-, dTTP dUTP, PCR-. -N-, 2 min 50OC.
    2. (, PCR-, ). , . , , .


     6.




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