Team:TUDelft/22 July 2009
From 2009.igem.org
22 July 2009
Sriram
Observed growth in all the 5 culture tubes [BBa_E0040 - GFP, BBa_C0040 - TetR, BBa_C0051 - cl, BBa_I12006 - pLamclin, BBa_E1010 - mRFP1] and at 11 AM the DNA extraction was started and done with miniprep plasmid DNA extraction kit. The DNA concentrations were noted for all the amplified biobrick parts using Nanodrop. [The values must be given below...It is in lab notes ... will be updated tomorrow...]
Agarose gel electrophoresis was run for all the 12 amplified biobrick parts with 2µl loading buffer for 20 minutes (17:30 to 17:50) and the gel is stored in fridge to continue with electrophoresis tomorrow.
Daniel
Prepared the glycerol stocks for the remaining culture of the 9 biobricks [BBa_E0040 - GFP, BBa_C0040 - TetR, BBa_C0051 - cl, BBa_I12006 - λp, BBa_E1010 - mRFP1 of Delay module and BBa_I714031 - OriT-R, BBa_J23100 - strong promoter, BBa_E0840 - GFP generator and BBa_I13522 - pTet GFP of Conjugation module] for future use. The protocol is as follows:
1. Centrifuge 1ml of culture in eppendorf at 8000rpm for 3 minutes at 15°C.
2. Decant the supernatant.
3. Add 1 ml of culture and centrifuge at 8000rpm for 3 minutes at 15°C.
4. Decant the supernatant.
5. Add 1 ml of culture and centrifuge at 8000rpm for 3 minutes at 15°C.
6. Decant the supernatant.
7. Add 1 ml of culture and centrifuge at 8000rpm for 3 minutes at 15°C.
8. Decant 0.5ml of the supernatant.
9. Resuspend the dense pellet throughly, add 0.5 ml of 10% glycerol and store in -80°C freezer.
Calin
Observed growth in all the 4 culture tubes [BBa_I714031 - OriT-R, BBa_J23100 - strong promoter, BBa_E0840 - GFP generator and BBa_I13522 - pTet GFP] and at 11 AM the DNA extraction was started and done with miniprep plasmid DNA extraction kit. The DNA concentrations were noted for all the amplified biobrick parts using Nanodrop.
Part | Concentration (ng/uL) |
BBa_I714031 OriT-R | 24.5 |
BBa_J23100 strong promoter | 56.5 |
BBa_E0840 GFP generator | 15.1 |
BBa_I13522 pTet GFP | 19.5 |
Agarose gel electrophoresis was run for all the 4 amplified biobrick parts, with 2µl loading buffer for 20 minutes (17:30 to 17:50)and the gel is stored in fridge to continue with electrophoresis tomorrow.