2014hs.igem.org/team:Charlottesville RS/Notebook/041814

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(→April 18th, 2014)
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''Alli Ambrosini''
''Alli Ambrosini''
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Did research on how we will be testing the results of our project in class today. We will need to get non-nutrient agar plates so that we can test whether the bacteria can eat our PHB at all. Normal agar has dextrose in it, which bacteria will consume before they touch PHB because its easier to eat. A simple sugar is a lot easier to break down than a long polymer such as PHB.  
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Did research on how to test the results of our project. The team will need to get non-nutrient agar plates so that it can be tested whether the bacteria can eat our PHB at all. Normal agar has dextrose in it, which bacteria will consume before they touch PHB because its easier to eat. A simple sugar is a lot easier to break down than a long polymer such as PHB.  
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''Lauren Ewell, John Grammer''
''Lauren Ewell, John Grammer''
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We took competent cells, centrifuged them to get a pellet. We resuspended in Luria broth, centrifuged and then we combined into three tubes and added calcium chloride and put on ice for 20 minutes. Then Alli took over after that step. She centrifuged gently by pulsing 10 times and then resuspended with 200uL of cold calcium chloride.
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Took competent cells, centrifuged them to get a pellet. Resuspended in Luria broth, centrifuged and then combined into three tubes and added calcium chloride and put on ice for 20 minutes. Alli centrifuged gently by pulsing 10 times and then resuspended with 200uL of cold calcium chloride.
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Did the re-suspending of DNA and transformation procedure.
Did the re-suspending of DNA and transformation procedure.
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1. With a pipette tip, punch a hole through the foil cover into the corresponding well of the Biobrick-standard part that you want. Make sure you have properly oriented the plate. We recommend that you do not remove the foil cover, as it could lead to cross contamination between the wells.
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The cells were possibly not competent because they were not suspended, there was a pellet at the bottom
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2. Pipette 10uL of dH2O (distilled water) into the well. Pipette up and down a few times and let sit for 5 minutes to make sure the dried DNA is fully re-suspended.
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The transformation procedure in the iGEM procedure notebook on page 28 was performed There were not 2 hours to incubate it, which was step 6
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3. Transform 5uL of the resuspended DNA into your competent cells, plate your transformation with the appropriate antibiotic* and grow overnight.
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Mrs. Minutella plated the transformed DNA at 60uL on each plate
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4. We are going to use the chloramphenicol plates. Be sure to label them.
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We weren’t sure the cells were competent because they weren’t suspended, there was a pellet at the bottom
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We then did the transformation procedure in the iGEM procedure notebook on page 28. We did not have 2 hours to incubate it, which was step 6
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[[https://2014hs.igem.org/Team:Charlottesville_RS/Notebook/Overview Back]]
[[https://2014hs.igem.org/Team:Charlottesville_RS/Notebook/Overview Back]]

Revision as of 13:24, 5 May 2014

April 18th, 2014

Alli Ambrosini

Did research on how to test the results of our project. The team will need to get non-nutrient agar plates so that it can be tested whether the bacteria can eat our PHB at all. Normal agar has dextrose in it, which bacteria will consume before they touch PHB because its easier to eat. A simple sugar is a lot easier to break down than a long polymer such as PHB.



Lauren Ewell, John Grammer

Took competent cells, centrifuged them to get a pellet. Resuspended in Luria broth, centrifuged and then combined into three tubes and added calcium chloride and put on ice for 20 minutes. Alli centrifuged gently by pulsing 10 times and then resuspended with 200uL of cold calcium chloride.



Alex Manchester, Nick Keen, Anders Beaurline, Jessica Prax, Bailey Fernandez

Did the re-suspending of DNA and transformation procedure.

The cells were possibly not competent because they were not suspended, there was a pellet at the bottom

The transformation procedure in the iGEM procedure notebook on page 28 was performed There were not 2 hours to incubate it, which was step 6

Mrs. Minutella plated the transformed DNA at 60uL on each plate

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