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Team:Wageningen UR/Journal/week2
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== Office work == | == Office work == | ||
| - | This is our first full week to work on | + | This is our first full week to work on our project. There are four of us who can work whole days (Robert, Jeroen, Kees and Mark) and two of us who can work in the afternoon (Thijs and Wouter), the others will join when the sixth period of our academic year ends . |
| - | + | This week we searched for programs which are able to predict the tertiary structure of the VLP monomers. Several of them were very interesting, but Phyre2 really stands out with the simple fill-in sheet and the possibility to download the PDB files of the predicted protein. Besides searching for programs, we also worked on the protocols that are going to be used and check if the required equipment and materials are available. Thijs and Mark worked also on the logo for the team and the template for the team-wiki. Jeroen and Wouter spent a day in the cold room to prepare large amounts of electrocompetent cells. | |
| - | This week we searched for programs which | + | |
| - | Jeroen and Wouter spent a day in the cold room to prepare large amounts of electrocompetent cells. | + | |
[Meeting] | [Meeting] | ||
Revision as of 08:36, 21 September 2012
week 2: 7 may - 13 may
Office work
This is our first full week to work on our project. There are four of us who can work whole days (Robert, Jeroen, Kees and Mark) and two of us who can work in the afternoon (Thijs and Wouter), the others will join when the sixth period of our academic year ends .
This week we searched for programs which are able to predict the tertiary structure of the VLP monomers. Several of them were very interesting, but Phyre2 really stands out with the simple fill-in sheet and the possibility to download the PDB files of the predicted protein. Besides searching for programs, we also worked on the protocols that are going to be used and check if the required equipment and materials are available. Thijs and Mark worked also on the logo for the team and the template for the team-wiki. Jeroen and Wouter spent a day in the cold room to prepare large amounts of electrocompetent cells.
[Meeting]
written by: Mark
Lab work
Growing cells
Monday:
- Checking plates
- All plates grown well accept for amp plates, due to drying effects.
- Growing E. coli in liquid LB. 3 plates per strain were picked and grown in 25 ml LB. All 3 strains in shaker, 180 RPM, 30 degree celcius.
- Grow them overnight
Tuesday:
- Continue growth.
- Culture mach1 and DH5X are plated 10x and 1000x diluted on LB plates. Dilutions are done with demi water. Plates were placed in 37 degree Celcius stove.
- Grow them overnight
Wednesday:
- Check plates
- 10x dilution was to low, overgrown.
- 1000x was nearly overrown, but still acceptable.
- Picked 1 MACH1 and 1 DH5X colony and grown in 10 ml SOB medium, 37 degree Celcius, 180 RPM
- Grow them overnight
Preparing competent cells
Thursday:
- Competent cells were prepared from both Mach1 and DH5-Alfa E. coli.
- following electrocompetent-cells protocol, with the following deviations:
- Starting culture: 4x 500 ml SOB
- Centrifuge rotor: SLA 3000
Testing CCMV protocol
Friday:
- Inoculated at 8:45
- IPTG at 11:45
- Centrifugation in greiner tubes
- Pellet stored at -20 degree Celcius, 50 ml greiner tubes
- Miniprep following the Gene Jet Fermentas protocol
- pet28
- standard 10 stock 1
- standard 10 stock 7
written by: Mark
