Paris/August 1

From 2007.igem.org

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(DGAT Assembly PCR: Megaprimer PCR)
(DGAT Assembly PCR: Megaprimer PCR)
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'''Quantity:''' 5µg/µL <br>
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'''Quantity:''' 5µg/µL
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'''Expected size:''' 1443 bp
'''Expected size:''' 1443 bp

Revision as of 14:20, 2 August 2007

DGAT Assembly PCR: Megaprimer PCR

  • Material:

- DNA plasmid (pKS::DGAT): 1.5µL (200ng)
- Tampon: 10µL
- dNTP: 1µL (final 200µmol each)
- Taq: 0.5µL
- Water: 29µL
- Primer: ForDGAT1: 4µL (final 0.8µM)
- Megaprimer: DGAT2: 4µL (50ng)

DGATDPst108012007.jpg
  • PCR Program:

5 cycles only with DGAT2 as primer:
- 98°C: 5min
- 98°C: 1min (X5)
- 60°C: 30s (X5)
- 72°C: 2min (X5)
- 72°C: 10 min
- 4°C: inf

Add primer ForDGAT1:

30 cycles: - 98°C: 5min
- 98°C: 1min (X5)
- 60°C: 30s (X5)
- 72°C: 2min (X5)
- 72°C: 10 min
- 4°C: inf


Quantity: 5µg/µL
Expected size: 1443 bp

  • After purification:

2µL of purification product (2µL of ladder)

DGATDPst108012007purification.jpg