From 2008.igem.org

Revision as of 12:07, 29 October 2008

Overview

First week

Monday 01/09/2008

• Preparation of TB media with an Agar concentration of 0.4 %, 0.2 % and 0.1 %.

First microscope test using MG1655 in 6 small chambers from Labtek:

3 types each type twice:
I - 0.1% agar in LB
II - 0.25% agar in LB
III - 0.5% agar in LB

View:

I II III
I II III

• In I and II each chamber was filled with just 100 µl LB agar to cover the whole ground.

• Because the III. solution was more viscous 150 µl were necessary to cover the ground.

• The bacteria were put into one corner of a chamber. (2 µl of a MG1655 overnight culture) They have been set under the media or in between.

• About that another 100 µl of a LB solution with 2 % agar were poured. After the second layer has solidified further 250 µl LB-media were added.

• The chambers were incubated overnight at 37 °C. For microscope on the next day.

Notes: Because of surface tensions the solutions tend to be thicker at the borders of the chamber than in the center.
-> In the further future 150 - 200 µl will be used.

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Tuesday 02/09/2008

First approach in microscope seems to be good and can be followed up.

Yesterday overnight cultures of E.Coli HCB33 and E.Coli MG1655 have been inoculated in 3 ml LB media. Today they were diluted with additional 4 ml LB and splitted into two 3 ml falcon-samples.
Followed by incubation at 37 °C.

Glycerol-stocks

• The other 1 ml of each sample was used to create a glycerol stock.
  
  • 1 ml of the ONC mixed with 150 µl of 80% glycerol by vortexing. Storage at -80 °C after 30 minutes of rest.

First test producing fluorescent strains HCB33 and MG1655 strains.

• Therefore our strains will be transformed with
  
  • 11tg a plasmid with Amp-resistance which should allow the expression of GFP.
    
  • BBa_I7100 from the registry, this should be expressing GFP, the production may be repressed with TetR.

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Wednesday 03/09/2008

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Thursday 04/09/2008

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Friday 05/09/2008

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Saturday 06/09/2008

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Sunday 07/09/2008

back up Overview 2nd Week

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