Team:Warsaw/Calendar-Main/11 May 2009

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PCR pho, mgtc, inv

Kama


Tasks:

  • Amplification of pho, mgtc and inv

Methods:

  • PCR mixture's composition:

    2,5μl pfu buffer (Fermentas) 
    2,5μl MgSO4 (Fermentas) 
    1,5μl primers 
    1μl dNTPs (10 mM) 
    1μl template
    0,5μl pfu turbo polymerase (KNGiE)

    The solution was topped up with H2O to 25μl.

  • PCR programs:
  • pho

     2min30s 95°C 
    (30s 95°C, 35s 56°C, 3min30s 72°C)x3
    (30s 95°C, 35s 61°C, 3min30s 72°C)x28
    10min 72°C
    ~ 7°C

    mgtc

     1min30s 95°C 
    (30s 95°C, 35s 48°C, 1min 72°C)x3
    (30s 95°C, 35s 58°C, 1min 72°C)x28
    10min 72°C
    ~ 7°C

    inv

     4min 95°C 
    (30s 95°C, 1min 53°C , 4min15s 72°C)x31
    10min 72°C
    ~ 7°C