IGEM:Groningen/Notebook/iGEM 2011/2011/09/12: Difference between revisions
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<br> Extension: 72°C for 4 min. | <br> Extension: 72°C for 4 min. | ||
<br> Final extension: 72°C for 10 min. | <br> Final extension: 72°C for 10 min. | ||
<br> Store infinite at 4°C | <br> Store infinite at 4°C | ||
<br> | <br> Finished at 17h, put samples on gel and purify the PCR products. | ||
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<br> Making scheme for digestion and ligation of the 31 samples: | |||
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<br> Digestion: | <br> Digestion: | ||
<br> Digestion will be done in one reaction (so insert and vector will be in one tube, so don't use fastAP!) | <br> Digestion will be done in one reaction (so insert and vector will be in one tube, so don't use fastAP!) | ||
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<br> 2μl FD buffer | <br> 2μl FD buffer | ||
<br> 9μl MQ water | <br> 9μl MQ water | ||
<br> | <br> | ||
<br> incubate samples for 1h at 37 degrees | <br> incubate samples for 1h at 37 degrees | ||
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<br> Ligate overnight at 4 °C | <br> Ligate overnight at 4 °C | ||
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<br> Purification of the plasmid backbone PCR product after finished PCR. | <br> Purification of the plasmid backbone PCR product after finished PCR at 17:15. | ||
<br> Analysed at 1% agarose gel: | <br> Analysed at 1% agarose gel: | ||
<br> Digestion and ligation was not done, since the PCR of the plasmid backbone failed. | <br> Digestion and ligation was not done, since the PCR of the plasmid backbone failed. |
Revision as of 10:10, 12 September 2011
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12-9-11
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