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Team:Bordeaux/NotebookAugust19
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Revision as of 01:48, 18 October 2014
19th August
Molecular biology
- Verification of our plasmid Miniprep we haven’t any problems with the plasmid
-
Digestion of 1μg of insert
Ex : ELP = 460ng/μl
So 1μg (ELP) = 2,17μlDNA 10μl
+ 7,83μl H2O
Xba1 0,5μl
Spe1 0,5μl
10X Buffer 4 5μl
H+O qsp 50μl
Migration on agarose 1,5% gel (100V, 30min)
-
Test of pSB1C3 miniprep
- Digestion : we want 1μg of plamisd
- Ligation : [insert] = 6 [plasmid] => molar concentration
- PCR: linearization of ELP-40
Molecular biology
- Problem with our 1x500ml Erlenmeyer bacteria grow slower than in 2x250ml erlen.
- Preculture of ELP-60 in 2x250ml of TB+amp
- Preculture of BL21 in 5ml of LB overnight
