IGEM:Groningen/Notebook/iGEM 2011/2011/08/01: Difference between revisions
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<br> | <br> | ||
<br>PBADaraC | <br>PBADaraC | ||
<br> insert: | <br> insert: 4μl | ||
<br> EcoRI: 1μl | <br> EcoRI: 1μl | ||
<br> SpeI: 1μl | <br> SpeI: 1μl | ||
<br> Fast digest buffer: 2μl | <br> Fast digest buffer: 2μl | ||
<br> MQ: | <br> MQ: 12μl | ||
<br> Digest for 1h at 37 degrees | <br> Digest for 1h at 37 degrees | ||
<br> DNA clean up | <br> DNA clean up | ||
Line 60: | Line 60: | ||
<br> PBADaraC-cI-LVA-DT | <br> PBADaraC-cI-LVA-DT | ||
<br> Vector: 8.5μl | <br> Vector: 8.5μl | ||
<br> insert: . | <br> insert: 6.1μl | ||
<br> T4 DNA ligase: 1μl | <br> T4 DNA ligase: 1μl | ||
<br> T4 DNA ligase buffer: 2μl | <br> T4 DNA ligase buffer: 2μl | ||
<br> MQ: . | <br> MQ: 2.4μl | ||
<br> | <br> | ||
<br> PBADaraC-LasR-LVA-DT | <br> PBADaraC-LasR-LVA-DT | ||
<br> Vector: 8.5μl | <br> Vector: 8.5μl | ||
<br> insert: . | <br> insert: 6.1μl | ||
<br> T4 DNA ligase: 1μl | <br> T4 DNA ligase: 1μl | ||
<br> T4 DNA ligase buffer: 2μl | <br> T4 DNA ligase buffer: 2μl | ||
<br> MQ: . | <br> MQ: 2.4μl | ||
<br> | <br> | ||
<br> PBADaraC-RBS-GFP-DT | <br> PBADaraC-RBS-GFP-DT | ||
<br> Vector: 8.5μl | <br> Vector: 8.5μl | ||
<br> insert: . | <br> insert: 6.1μl | ||
<br> T4 DNA ligase: 1μl | <br> T4 DNA ligase: 1μl | ||
<br> T4 DNA ligase buffer: 2μl | <br> T4 DNA ligase buffer: 2μl | ||
<br> MQ: ... | <br> MQ: 2.4μl | ||
<br> | |||
<br> Selfligation tests: | |||
<br> Vector: 8.5μl | |||
<br> T4 DNA ligase: 1μl | |||
<br> T4 DNA ligase buffer: 2μl | |||
<br> MQ: 8.5μl | |||
<br> | <br> | ||
<br> Ligate for 30 min at room temperature | <br> Ligate for 30 min at room temperature |
Revision as of 07:51, 1 August 2011
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1-8-11
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