User:Nicole Bonan/Notebook/Chem 571 Lab Notebook/2015/09/29: Difference between revisions
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# We then incubated both samples in a 37 degree C water bath for 1 hour | # We then incubated both samples in a 37 degree C water bath for 1 hour | ||
# After that, we began making our standards. Our standard samples were made as follows: | # After that, we began making our standards. Our standard samples were made as follows: | ||
## Standard A: 150uL of the reaction sample from step | ## Standard A: 150uL of the reaction sample from step 3 | ||
## Standard B: 75uL of Standard A and 75uL of water | ## Standard B: 75uL of Standard A and 75uL of water | ||
## Standard C: 75uL of Standard B and 75uL of water | ## Standard C: 75uL of Standard B and 75uL of water | ||
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## Standard G: 75uL of Standard F and 75uL of water | ## Standard G: 75uL of Standard F and 75uL of water | ||
## Standard H: 75uL of Standard G and 75uL of water | ## Standard H: 75uL of Standard G and 75uL of water | ||
# We then made our blanks as follows: | |||
## Blank A: 150uL of the reaction sample from step 4 | |||
## Blank B: 75uL of Blank A and 75uL of water | |||
## Blank C: 75uL of Blank B and 75uL of water | |||
## Blank D: 75uL of Blank C and 75uL of water | |||
## Blank E: 75uL of Blank D and 75uL of water | |||
## Blank F: 75uL of Blank E and 75uL of water | |||
## Blank G: 75uL of Blank F and 75uL of water | |||
## Blank H: 75uL of Blank G and 75uL of water | |||
# In separate 1.5mL Eppindorf tubes, we then combined the following (for each standard) just before we took its fluorescence measurements: | # In separate 1.5mL Eppindorf tubes, we then combined the following (for each standard) just before we took its fluorescence measurements: | ||
## 20uL of a standard | ## 20uL of a standard |
Revision as of 21:17, 3 October 2015
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ObjectiveFirst, we'll make a standard curve for the fluorescence of our proteinase, alpha-chymotrypsin. Then, we'll measure the fluorescence of samples of protease + lysozyme and use our standard curve in order to determine how the proteinase degrades the lysozyme. ProtocolDr. Hartings's protocol for today is in his lab notebook. Today, my group was doing the fluorescence analysis of protein degradation. Our protocol was as follows:
DataSee tomorrow's lab entry, as we did not have enough data today to analyze correctly. |