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Team:Warsaw/Calendar-Main/20 July 2009
From 2009.igem.org
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| - | *Accidentally I used the whole DNA samples extracted on Thursday to perform the digestion. I Added the digestion mix to my samples, hence the wrong buffer concentration and improper digestion on Friday. Thinking saves time and money. But there is another reason to justify the plasmid extraction. Now I know that | + | *Accidentally I used the whole DNA samples extracted on Thursday to perform the digestion. I Added the digestion mix to my samples, hence the wrong buffer concentration and improper digestion on Friday. Thinking saves time and money. But there is another reason to justify the repeated plasmid extraction. Now I know that my transformants after all have the correct plasmid so I can do the alkaline lysis on very few samples and use the AA kit to get greater DNA concentration. After all this is my insert for the next ligation, so we need a lot. I know that no one will ever read the notebook up to this point. |
Revision as of 10:32, 22 July 2009
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Insertion of the pho gene into the pKSII+ plasmid
Kama
The ligation mix was prepared as follows: 2μl plasmid, 8μl gene, 3μl ligation buffer, 1 μl T4 DNA ligase (Fermentas), 0,5μl PEG the solution was topped up with H2O to the final volume of 30 μl. As a control:2μl plasmid, 3μl ligation buffer, 1 μl T4 DNA ligase (Fermentas), 0,5μl PEG the solution was topped up with H2O to the final volume of 30 μl. The ligation was carried out in 18°C overnight (~15h)
Construction of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012 K177012] operon1_part2
Ania
Tasks:
- Set up a liquid culture from transformants containing [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0032 BBa_B0032 - RBS.3] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 BBa_C0012 - lacI repressor] on the pSB1A2 ampicillin resistant plasmid in order to amplify the plasmid and extract it from the bacteria.
Comment:
- Accidentally I used the whole DNA samples extracted on Thursday to perform the digestion. I Added the digestion mix to my samples, hence the wrong buffer concentration and improper digestion on Friday. Thinking saves time and money. But there is another reason to justify the repeated plasmid extraction. Now I know that my transformants after all have the correct plasmid so I can do the alkaline lysis on very few samples and use the AA kit to get greater DNA concentration. After all this is my insert for the next ligation, so we need a lot. I know that no one will ever read the notebook up to this point.
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