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Team:Newcastle/24 August 2010
From 2010.igem.org
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Contents |
Miniprep for pGFPrrnB with filamentous cell part
Aims
The aim of this experiment is to obtain stock of the filamentous cell part in pGFP-rrnB for transformation into Bacillus subtilis 168.
Materials and Protocol
Please refer to qiagen minipreps and nanodrop spectrophotometer protocols.
Results
| Tube 1 | Tube 2 | Tube 3 | Tube 4 | Tube 5 | Tube 6 | Tube 7 | Tube 8 | Tube 9 | Tube 10 | Tube 11 | Tube 12 |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 286.1 µl/ml | 304.0 µl/ml | 316.3 µl/ml | 421.6 µl/ml | 518.7 µl/ml | 460.1 µl/ml | 370.1 µl/ml | 377.3 µl/ml | 346.0 µl/ml | 347.4 µl/ml | 202.8 µl/ml | 307.4 µl/ml |
Table 1: Nanodrop spectrophotometer results. Table represents the amount of plasmid present in µl/ml quantity.
Discussion
The concentration ranged from 286.1 µl/ml to 518.7 µl/ml. Therefore we have obtained high concentration of yneA in pGFPrrnB.
Single and Double Digestion of pGFPrrnB with filamentous cell part
Aims
The aim of this experiment is screen our minipreps to check if the insert is present.
Materials and Protocol
We are doing two digests for pGFPrrnB and yneA:
- Single digest with HindIII;
- Double digest with EcoRI and NheI.
Please refer to restriction digests and gel electrophoresis.
