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Team:Stockholm/1 October 2010
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I inoculated 12 ml LB and 24 ul chloramphenicol with IgG protease_Tra10_Ntermin#6 again since I acidentally droped the previous sample and therefore lost it. | I inoculated 12 ml LB and 24 ul chloramphenicol with IgG protease_Tra10_Ntermin#6 again since I acidentally droped the previous sample and therefore lost it. | ||
| - | ===iGEM Uppsala collaboration== | + | ===iGEM Uppsala collaboration=== |
I drove to Uppsala and started a collaboration with their team. I obtained a construct that they want me/our group to PCR with verification primers (VF2 & VR) and run on an agarose gel. I in turn gave them the tyrosinase gene to also amplify via PCR but with own designed primers. | I drove to Uppsala and started a collaboration with their team. I obtained a construct that they want me/our group to PCR with verification primers (VF2 & VR) and run on an agarose gel. I in turn gave them the tyrosinase gene to also amplify via PCR but with own designed primers. | ||
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Andreas
LB agar plates
- 20 x 100 μg/ml Amp LB agar plates
Assembly of His⋅SOD⋅cCPP constructs
Digested pMA.His⋅SOD constructs for later assembly into cCPP plasmids, digested by Johan.
Digestion
| pMA.His⋅ SOD | |
|---|---|
| 10X FastDigest buffer | 3 |
| DNA (3 μg) | 7.5 |
| dH2O | 17.5 |
| FD EcoRI | 1 |
| FD AgeI | 1 |
| 30 μl |
- Incubation: 37 °C, 0:30
- Inactivation: 80 °C, 20 min
Stored in -20 °C for later ligation.
Nina
Miniprep
I performed a miniprep on IgG protease_Tra10_Ntermin#4 and Fusion_NS#2 according to the procedure that is described in protocols.
Send for sequencing
I sent two samples for seqeuncing. The mixture was 15 ul sample and 1.5 ul forward bank vector verification primer VF.
- IgG protease_Tra10_Ntermin#4 ASB0045 682
- Fusion_NS#2 ASB0045 681
Overnight culture
I inoculated 12 ml LB and 24 ul chloramphenicol with IgG protease_Tra10_Ntermin#6 again since I acidentally droped the previous sample and therefore lost it.
iGEM Uppsala collaboration
I drove to Uppsala and started a collaboration with their team. I obtained a construct that they want me/our group to PCR with verification primers (VF2 & VR) and run on an agarose gel. I in turn gave them the tyrosinase gene to also amplify via PCR but with own designed primers.
- Uppsala iGEM team's PCR master mix and prgm:
