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Team:Bordeaux/NotebookJune24
From 2014.igem.org
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<li>Test <a href="https://docs.google.com/spreadsheets/d/1_1G_XSVk9BdI5JrVnN87MqkYWdlEvxf_-ecaP0UAxfk/edit?usp=sharing">preculture</a> 200ml | <li>Test <a href="https://docs.google.com/spreadsheets/d/1_1G_XSVk9BdI5JrVnN87MqkYWdlEvxf_-ecaP0UAxfk/edit?usp=sharing">preculture</a> 200ml | ||
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13h of incubation for 20 et 40 (Glycerol)<br> | 13h of incubation for 20 et 40 (Glycerol)<br> | ||
Latest revision as of 00:32, 18 October 2014
24th June
Molecular biology
- Ligation pSB6A1 and VPGXG-20 of 23/06/14
2µl of digestion product
1,5µl insert - ratio 1 vector/3 insert
1µl Buffer ligase 10X
0,5µl T4 ligase
H2O qps 10µl
30 min at 16°C - Digestion ELP-40 and -60 and vector pSB1C3
Production
- Test preculture 200ml
13h of incubation for 20 et 40 (Glycerol)
13h of incubation for 60 (glucose)
For the next preculture : 200ml / 13h of incubation / put less bacteria in the preculture medium.
- Seeding at 11h
We want OD = 0,12 in 1L
C1V1=C2V2
V1 (preculture 20) = 0,12/2,777=43,2ml of preculture medium in 1L LB+Glycerol
V (préculture 40) = 0,12/2,609=46ml -> 40/glycérol
V (préculture 60) = 0,12/2,853=42ml -> 60/glycérol
DO T=0 (11h10)
20: 0,164
40: 0,093
60: 0,160
T=9h30 (17h40)
20: 1,802
40: 1,713
60: 1,508
