Team:Newcastle/Labwork/13 August 2009

=Formal Lab Session - 13th August 2009=

= Overview = 
 * Sporulation Tuning/Chassis Team - attempted to transform Bacillus subtilis cells with pGFP-rrnB as practice

Summary
Yesterday's germination attempt seems to have worked, but results are not yet clear. This may have been due to a calculation error for the lysozyme, where instead of adding 40ul of stock lysozyme per ml of buffer solution, 4ul of stock lysozyme was added instead.

Therefore, on Monday, we intend to redo the experiment. See Monday, 17th August for more details.

Today we are trying to transform Bacillus subtilis with gfp-rrnb integration vector using the Transformation Protocol as well as following the changes which the Metal Sensor Team implemented. Metal sensor team kindly inoculated B. subtilis cells into flask tubes and placed them into the shaking incubator. The overnight cultures were labelled 1, 2 and 3, while the control was labelled as 4. Our team used overnight culture tube 3 and control tube 4.

Results
Following an which clearly explains what to plate out from the Bacillus Transformation Protocol, we plated 4 plates, 3 of which contained the antibiotic chloramphenicol, and LB, and 1 containing just LB. The plate which contained just LB was the control plate.

The following pictures show the results of our team's transformations, which were a success.



No colonies were expected to grow on this plate as no DNA was added to the cells for transformation



Instead of adding DNA, water was added instead. Therefore, no colonies should be growing on this plate either.

Looking at the two pictures above, it shows the transformation has been a success as colonies were growing on the plates where DNA was added. The plate where 200ul of the transformed bacteria were added, had more colonies growing as expected.