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Team:Newcastle/Labwork/13 August 2009
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| + | __NOTOC__ | ||
| + | [[Image:Team Newcastle 2009 iGEM ProbationaryP-Sign.PNG|50px|right]] | ||
| + | =Formal Lab Session - 13th August 2009= | ||
| - | = | + | =<font color="Orange"><u>Overview</u></font>= |
| + | <font color="Orange"> | ||
| + | *[[#Sporulation Tuning/Chassis Team|Sporulation Tuning/Chassis Team]] '''- attempted to transform ''Bacillus subtilis'' cells with ''pGFP-rrnB'' as practice''' | ||
| + | </font> | ||
| + | <br> | ||
==<u>Sporulation Tuning/Chassis Team</u>== | ==<u>Sporulation Tuning/Chassis Team</u>== | ||
| - | [https://2009.igem.org/ | + | ===Summary=== |
| + | [[Image:Newcastle 13 August BSub Transformation test plates.jpg|thumb|The plates for our test transformation.]] | ||
| + | [https://2009.igem.org/Team:Newcastle/Labwork/12_August_2009#Sporulation_Tuning.2FChassis_Team 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 [https://2009.igem.org/ | + | Therefore, on [https://2009.igem.org/Team:Newcastle/Labwork/17_August_2009#Sporulation_Tuning.2F_Chassis_Team Monday], we intend to redo the experiment. See [https://2009.igem.org/Team:Newcastle/Labwork/17_August_2009#Sporulation_Tuning.2F_Chassis_Team Monday, 17th August] for more details. |
| - | Today we are trying to transform ''Bacillus subtilis'' with gfp-rrnb integration vector using the [https://2009.igem.org/Team:Newcastle/Project/Labwork/OurProtocols/TransformBac Transformation Protocol]. 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. | + | Today we are trying to transform ''Bacillus subtilis'' with gfp-rrnb integration vector using the [https://2009.igem.org/Team:Newcastle/Project/Labwork/OurProtocols/TransformBac Transformation Protocol] as well as following the [https://2009.igem.org/Team:Newcastle/Labwork/11_August_2009#Changes_to_the_Bacillus_transformation_protocol_.28day_of_transformation.29 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 [https://2009.igem.org/Image:Team_Newcastle_2009_Bacillus_Transformation_Protocol_Steps7-9.PNG illustration] which clearly explains what to plate out from the [https://2009.igem.org/Team:Newcastle/Project/Labwork/OurProtocols/TransformBac 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. | ||
| + | |||
| + | [[Image:Newcastle 14 August BSub Transformation test control.jpg|thumb|250px|left|The transformation control plate.]] | ||
| + | |||
| + | No colonies were expected to grow on this plate as no DNA was added to the cells for transformation | ||
| + | |||
| + | [[Image:Newcastle 14 August BSub Transformation test h2o.jpg|thumb|250px|left|The transformation control (water) plate.]] | ||
| + | |||
| + | Instead of adding DNA, water was added instead. Therefore, no colonies should be growing on this plate either. | ||
| + | |||
| + | |||
| + | [[Image:Newcastle 14 August BSub Transformation test 50.jpg|250px|The transformation 50ul plate.]] | ||
| + | [[Image:Newcastle 14 August BSub Transformation test 200.jpg|250px|The transformation 200ul plate.]] | ||
| + | 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. | ||
| + | |||
| + | {{:Team:Newcastle/Project/Labwork/CalTemplate}} | ||
{{:Team:Newcastle/Footer}} | {{:Team:Newcastle/Footer}} | ||
{{:Team:Newcastle/Right}} | {{:Team:Newcastle/Right}} | ||
Latest revision as of 14:19, 20 October 2009
Project
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Links
Formal Lab Session - 13th August 2009
Overview
- Sporulation Tuning/Chassis Team - attempted to transform Bacillus subtilis cells with pGFP-rrnB as practice
Sporulation Tuning/Chassis Team
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 illustration 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.
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News
Events
- 20 – 21 June 2009 - Europe workshop (London)
- 23 – 24 June 2009 - UK iGEM meetup (Edinburgh)
- 23 October Practice Presentation (Newcastle)
- 23 October T-shirts are ready
- 27 October Practice Presentation (Sunderland)
- 27 October Poster is ready
- 30 October – 2 November 2009 - Jamboree (Boston)
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