Team:TU Munich/lab/notebook/solidmatrix

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Preliminary Experiment 1: The Search For The Perfect Solid Medium (most work done by Thorsten, Anna, Simon)

04-11-2011

light transmission of solid agar, different agar concentrations and growth media

Purpose:

In order to create the 3D-Printer based on a double-light-sensitive-promotor, it is not only necessary to immobilize the cells within a solid medium but also to be able to induce the protein expression at a definied spot. Thus, we want to examine the light transmission properties of different solid media.

Procedure:

To gain data on the light transmission of different solid media, different compositions of media were produced and poured into 1 ml cuvettes while they were still liquid. After gelation, a transmission spectrum (230 nm - 750 nm) was recorded for each sample


Agar with LB-Medium:

Agar-concentration [w/v] Agar-Agar [g] LB-Medium [ml] number of cuvettes
0.5 %0.05103
1.0 %0.10103
1.5 %0.15103
2.0 %0.20103
2.5 %0.25103
3.0 %0.30103

Agar with M9-Medium:

Agar-concentration [w/v] Agar-Agar [g] M9-Medium 5x [ml] distilled water [ml] number of cuvettes
0.5 %0.05284
1.0 %0.10283
1.5 %0.15283
2.0 %0.20283
2.5 %0.25283
3.0 %0.30283

Agarose with M9-Medium:

Agarose-concentration [w/v] Agarose [g] M9-Medium 5x [ml] distilled water [ml] number of cuvettes
0.5 %0.05284
3.0 %0.30283


Results & Discussion:

The transmission spectra were saved locally on the lab-computer (upload?)

LB-medium has low tranmission values at shorter wavelengths (hence its yellow appearance) and should therefore be avoided in our experiment if possible. If a clear medium (such as M9) is sufficient for our purposes it should be chosen. It has no significant absorbtion of blue light, which makes the application of a promotor sensitive to blue light feasible.

Gels prepared with Agarose were clearer than those prepared with Agar-Agar.

The transmission values are higher at lower concentrations of the gelling agent.


Is a glas or plastic container neccessary for solid media?

Purpose:

We figured that if we could do without a glas or plastic container surrounding the solid medium, this would increase the precision of our light-beam aiming at the cells that are to be induced (no light refraction at the container walls, etc.).

Procedure:

To examine if it is possible to make a detachted, self-supporting block of agar, we made a 40 ml block of 0.5 % (w/v) Agar with M9-Medium in a small beaker glass. After the gellation process we turned it out and left it to stand over night.

Results & Discussion:

The block had lost a lot of water over night, so that it was standing in a puddle. Therefore it's not possible to make a detachted, self-supporting block of agar (0.5 % w/v).