User:Beatriz Gimenez De C./Notebook/571/2014/09/10: Difference between revisions
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*Make 2 duplicate blanks (1 mL Tris/NaCl bufferand 200 μL BRADFORD reagent + 800 μL buffer) | *Make 2 duplicate blanks (1 mL Tris/NaCl bufferand 200 μL BRADFORD reagent + 800 μL buffer) | ||
*repeat the process using Lysozyme. | *repeat the process using Lysozyme. | ||
==Results== | |||
* UV-Vis data from the 6 serial dilutions of '''BSA''' (1 μg/mL, 2 μg/mL, 5 μg/mL, 10 μg/mL, 15 μg/mL, 20 μg/mL) | |||
* personal note : Graph and table yet to be uplaoded ( need to ask how it is done) | |||
*UV-Vis data from the 6 serial dilutions of '''Lysozyme''' (1 μg/mL, 2 μg/mL, 5 μg/mL, 10 μg/mL, 15 μg/mL, 20 μg/mL) | |||
* personal note : Graph and table yet to be uplaoded ( need to ask how it is done) | |||
==Calculations== | |||
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Revision as of 09:56, 18 September 2014
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Task
- Prepare 50 mL of a standard saline solution (0.9 wt-% NaCl). Store in a 45 mL Falcon tube.
- Prepare 50 mL of a 50 mM Tris (not Tris-HCl) 50 mM NaCl solution. Store in a 45 mL Falcon tube.
- Prepare a stock solution of BSA that is roughly 5 mg in 5 mL of saline.
- Record UV-VIS spectra between 200 nm and 800 nm.
- 6 - 8 standard solutions 1 μg/mL and 20 μg/mL. Using determined volume of stock solution to add to 1.5 mL centrifuge tube. Add 200 μL of the Bio-Rad Protein Assay reagent. Use 1:4 concentrate diluted with water. Add to the solution Tris/NaCl until 1 mL volume is reached.
- vortex tubes and let them sit for 5 min.
- Obtain a UV-VIS spectrum (between 400 and 800 nm).
- Make 2 duplicate blanks (1 mL Tris/NaCl bufferand 200 μL BRADFORD reagent + 800 μL buffer)
- repeat the process using Lysozyme.
Results
- UV-Vis data from the 6 serial dilutions of BSA (1 μg/mL, 2 μg/mL, 5 μg/mL, 10 μg/mL, 15 μg/mL, 20 μg/mL)
- personal note : Graph and table yet to be uplaoded ( need to ask how it is done)
- UV-Vis data from the 6 serial dilutions of Lysozyme (1 μg/mL, 2 μg/mL, 5 μg/mL, 10 μg/mL, 15 μg/mL, 20 μg/mL)
- personal note : Graph and table yet to be uplaoded ( need to ask how it is done)