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Team:Stockholm/30 June 2010
From 2010.igem.org
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| - | == Expression of SOD and yCCS from pEX expression vector == | + | === Expression of SOD and yCCS from pEX expression vector === |
''Continued from 29/6'' | ''Continued from 29/6'' | ||
We realized that Top10 and DH5alpha are not suitable for IPTG-induced protein expression. SOD and yCCS expression was therefore not proceeded from ON cultures. '''Glycerol stocks''' were prepared from ON culture and '''pEX.SOD''' and '''pEX.yCCS plasmids''' were prepared. | We realized that Top10 and DH5alpha are not suitable for IPTG-induced protein expression. SOD and yCCS expression was therefore not proceeded from ON cultures. '''Glycerol stocks''' were prepared from ON culture and '''pEX.SOD''' and '''pEX.yCCS plasmids''' were prepared. | ||
| - | + | '''DNA concentrations''' | |
| - | + | {|border="1" cellspacing="0" cellpadding="2" | |
| - | |Sample | + | |'''Sample''' |
| - | | | + | |'''DNA conc (ng/ul)''' |
| - | |A(260)/A(280) | + | |'''A(260)/A(280)''' |
|- | |- | ||
|pEX.SOD | |pEX.SOD | ||
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|} | |} | ||
| - | + | === Preparation of competent BL21(DE3) cells === | |
| - | + | * 5 ml LB was inoculated with BL21(DE3) cells and grown ON in 37°C, 250 rpm. | |
| - | == Preparation of competent BL21(DE3) cells == | + | |
| - | * | + | |
Revision as of 10:03, 6 July 2010
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Contents |
Morning meeting
We discussed the project proceedings.
- It was decided that already amplified genes (cloned in pEX vector) should be transferred to pSB1C3 vector.
- IgG protease
- Superoxidase dismutase (SOD)
- yCCS
- bFGF
- Primers for not-yet amplified genes should be redesigned to include the complete Assembly standard 25 prefix and suffix.
- CPP
- Transportan 10
- LMWP
- TAT
- MITF
- Protein A Z-domain
- Tyrosinase
- Vitamin B9 genes
- CPP
- BL21(DE3) was chosen as our strain for IPTG-induced protein expression from pEX vector.
Andreas
Expression of SOD and yCCS from pEX expression vector
Continued from 29/6
We realized that Top10 and DH5alpha are not suitable for IPTG-induced protein expression. SOD and yCCS expression was therefore not proceeded from ON cultures. Glycerol stocks were prepared from ON culture and pEX.SOD and pEX.yCCS plasmids were prepared.
DNA concentrations
| Sample | DNA conc (ng/ul) | A(260)/A(280) |
| pEX.SOD | 18.15 | 1.98 |
| pEX.yCCS | 20.55 | 1.97 |
Preparation of competent BL21(DE3) cells
- 5 ml LB was inoculated with BL21(DE3) cells and grown ON in 37°C, 250 rpm.
