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Team:Newcastle/Labwork/27 July 2009
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Formal Lab Session - 27th July 2009
Running the gel for RFP and GFP bioricks
Today we prepared the gel with 0.8% agorose for last week's rescued plasmids which contain RFP and GFP biobricks. The final solution was 500ml using 4 gr of agorose in total.
We mixed 9ul of DNas with 1ul of the buffer and centrifuged shortly. The rest of the DNAs are then put back to the freezer.
Finally we run the gel. We loaded the ladder to the first and fourth wells, GFP to the 2nd and RFP to the 3rd well and set the voltage to 120V which was then reduced to 90V.
Below are the images:
Transformation of new biobricks
We prepared the agar plates and left the tubes including the biobricks for incubation overnight.
We checked the agar plates containing colonies of plasmids with five different biobricks from last week. We will be following Phil’s mini method for Alkaline Lysis for Mini Prep The list of plates and the biobricks are below:
| Plate | BioBrick | Number used for the labelling | Name |
| Plate 1 | BBa_C0056 | 1 | CI Coding Sequence |
| Plate 2 | BBa_B1002 | 2 | Terminator |
| Plate 3 | BBa_C0077 | 3 | CinR Coding sequence |
| Plate 4 | BBa_C0076 | 4 | CinI Coding Sequence |
| Plate 5 | BBa_R0077 | 5 | CinR sensitive promoter |
Except from plate 3, we had colonies formed in all plates.
Colonies from the plates were picked and mixed in a tube containing 5ml of LB. We prepared 12 tubes for 4 different biobricks. For each biobrick we labelled the tubes as A, B, and C. Finally we left them at shaking incubator overnight.
Preparation of the plates
We prepared the agar solutions and poured into the plates which are then placed into the fridge. The list of plates and their content are as below.
| Content | Number of plates prepared | Labelled as |
| LB | 4 | LB 27/07/09 |
| LB + Amp | 6 | LB+A 27/07/09 |
| LB + Amp + Kan | 5 | LB+AK 27/07/09 |
Tomorrow
We have placed the required solutions to the bench for the experiment apart from RNAase. We need to find it first thing in the morning tomorrow then we will follow Phil’s mini method for Alkaline Lysis for Mini Prep. GFP and RFP are placed into the freezer. The DNA ladder is in the fridge. Plates are placed into the fridge just below the top shelf. They were split into two since they did not all fit to the shelf.
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News
Events
- 20 – 21 June 2009 - Europe workshop (London)
- 23 – 24 June 2009 - UK iGEM meetup (Edinburgh)
- 23 October Practice Presentation (Newcastle)
- 23 October T-shirts are ready
- 27 October Practice Presentation (Sunderland)
- 27 October Poster is ready
- 30 October – 2 November 2009 - Jamboree (Boston)
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