IGEM:PennState/Labbook/NoahJohnson/2007-7-23: Difference between revisions
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*PCR End: 2:35 PM | *PCR End: 2:35 PM | ||
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Revision as of 07:21, 24 July 2007
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) 1741015
- 2) 1741017
- 3) 1741018
- 4) 1741019
- 5) 1741020
- 6) 1741021
- 7) 1741011
- 8) 1741023
- 9) 1741005
- 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 ∞
- Cycle 1: (1x)
- PCR Start: 1:09 PM
- PCR End: 2:35 PM
9:30-5
