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Team:Stockholm/30 June 2010
From 2010.igem.org
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(New page: {{Stockholm/Top2}} = Morning meeting = We discussed the project proceedings. * It was decided that already amplified genes (cloned in pEX vector) should be transferred to [http://partsre...) |
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''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 | + | 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. |
== Preparation of competent BL21(DE3) cells == | == Preparation of competent BL21(DE3) cells == | ||
* ON culture was set and grown in 37°C ON with 250 rpm rotary shaking. | * ON culture was set and grown in 37°C ON with 250 rpm rotary shaking. | ||
Revision as of 22:16, 1 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 [http://partsregistry.org/Part:pSB1C3 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.
Preparation of competent BL21(DE3) cells
- ON culture was set and grown in 37°C ON with 250 rpm rotary shaking.
