IGEM:PennState/Labbook/NoahJohnson/2007-7-23

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July 24, 2007

  • Added to each tube:
    • 5uL 10X Pfx Amplification buffer/ ThermoPol
    • 1.5uL 10mM dNTP
    • 1uL 50mM MgSO4
    • 1.5uL Primer Mix (0.75uL each)
    • 1uL Template DNA (10pg-200ng)
    • 0.5uL Pfx DNA Polymerase/ TAq Polymerase
    • 39.5uL Autoclaved dH2O (Enough to make 50uL total)
  • Template DNA Used:
    • 1) I741015
    • 2) I741017
    • 3) I741018
    • 4) I741019
    • 5) I741020
    • 6) I741021
    • 7) I741011
    • 8) I741023
    • 9) I741005
    • 10) (-)
  • Primers:
    • 1) 3398 & 3399
    • 2) 3400 & 3401
    • 3) 3406 & 3407
    • 4) 3408 & 3409
    • 5) 3402 & 3403
    • 6) 3404 & 3405
    • 7) 3412 & 3413
    • 8) 3410 & 3411
    • 9) 3416 & 3417
    • 10) 3410 & 3411
  • Thermocycler Protocol (pfx)
    • Cycle 1: (1x)
      • Step 1: 94°C for 2:00
    • Cycle 2: (28x)
      • Step 1: 94°C for 0:15
      • Step 2: 54°C for 3:00
      • Step 3: Gradient for 1:00
      • H . . G . . . F . . E . . . D . . C . . . B . . A
      • 60° 60.8° 61.9° 63.5° 65.8° 67.5° 68.5° 69°
    • Cycle 3: (1x) Hold at 4°C for ∞
  • PCR Start: 1:09 PM
  • PCR End: 2:35 PM

9:30-5

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