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

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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

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