IGEM:Groningen/Notebook/iGEM 2011/2011/06/21

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(Autocreate 2011/06/21 Entry for IGEM:Groningen/Notebook/iGEM_2011)
Current revision (05:25, 21 September 2011) (view source)
(Entry title)
 
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==Entry title==
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==21-6-11==
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* Insert content here...
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<br>
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<br> Digestion+ ligation PhybB product with taq polymerase
 +
<br>
 +
<br> Purify the PCR product with the High Pure PCR Purification kit from Roche
 +
<br>
 +
<br> Digestion mix:
 +
<br> 18μl buffer
 +
<br> 4.5μl EcoRI
 +
<br> 4.5μl SpeI
 +
<br> 49μl MQ water
 +
<br>
 +
<br> Vector mix:
 +
<br> 18μl buffer
 +
<br> 4.5μl EcoRI
 +
<br> 4.5μl XbaI
 +
<br> 49μl MQ water
 +
<br>
 +
<br> Ligation:
 +
<br> 2μl T4 DNA ligase buffer
 +
<br> 1μl T4 DNA ligase
 +
<br> 7μl insert
 +
<br> 3μl vector
 +
<br> 7μl MQ water
 +
<br>
 +
<br> Ligate 2 hours at room temperature
 +
<br> Ligate further overnight
 +
<br>
 +
<br> Helping with sponsoring
 +
<br> Making researchplan
 +
<br>
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21-6-11



Digestion+ ligation PhybB product with taq polymerase

Purify the PCR product with the High Pure PCR Purification kit from Roche

Digestion mix:
18μl buffer
4.5μl EcoRI
4.5μl SpeI
49μl MQ water

Vector mix:
18μl buffer
4.5μl EcoRI
4.5μl XbaI
49μl MQ water

Ligation:
2μl T4 DNA ligase buffer
1μl T4 DNA ligase
7μl insert
3μl vector
7μl MQ water

Ligate 2 hours at room temperature
Ligate further overnight

Helping with sponsoring
Making researchplan

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