Paris/August 1

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(DGAT Assembly PCR: Megaprimer PCR)
 
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[[Paris/July 31|yesterday]] -- [[Paris/August 2|tomorrow]] <br>
== DGAT Assembly PCR: Megaprimer PCR ==
== DGAT Assembly PCR: Megaprimer PCR ==
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- Megaprimer: DGAT2: 4µL (50ng)<br>
- Megaprimer: DGAT2: 4µL (50ng)<br>
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[[Image: DGATDPst108012007.jpg|right]]
+
[[Image: DGATDPst108012007.jpg|right|100px]]
* PCR Program:<br>
* PCR Program:<br>
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- 4°C: inf
- 4°C: inf
-
<br>
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* '''Quantity:''' 5ng/µL <br>
-
 
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* '''Expected size:''' 1443 bp
-
'''Quantity:''' 5µg/µL
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* After purification: <br>
-
'''Expected size:''' 1443 bp
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[[Image: DGATDPst108012007purification.jpg|right|100px]]
 +
2µL of purification product (2µL of ladder): <br>
 +
- well 0: ladder<br>
 +
- well 1: negative control of purification<br>
 +
- well 2: purification product<br>

Latest revision as of 17:52, 7 October 2007

yesterday -- tomorrow

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: 5ng/µL
  • Expected size: 1443 bp
  • After purification:
DGATDPst108012007purification.jpg

2µL of purification product (2µL of ladder):
- well 0: ladder
- well 1: negative control of purification
- well 2: purification product