Team:TUDelft/22 July 2009

From 2009.igem.org

Lab Notebook

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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.

Part Concentration (ng/µl)
BBa_E1010 mRFPI 22.2
BBa_C0051 cI 19.4
BBa_C0040 TetR 19.4
BBa_ E0040 GFP 30.6
BBa_I12006 pLambclin 33
BBa_J23008 key3c 28.7
BBa_B0034 RBS 29.4
BBa_J23031 lock3c 23.3
BBa_B0015 Double Terminator 45.7
BBa_R0040 pTet 19.5
BBa_R0010 pLacI 25.8
BBa_K081013 RBS-cI-RBS 38.7

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.

Tim Weenink

Observed colonies on the plates from yesterdays transformation.


Weenink first gel.png