User:Monika Gasiorek/Notebook/CHEM-571 2014F/2014/10/15

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October 15, 2014

Tornado Warning

Disruptive...

SDS-PAGE: Running of the Gels

The four samples prepared yesterday were heated to 40C on a heating block. Afterwards, they were transferred into a premade gel along with a premade marker/ladder. The gel was then run under the same conditions as done earlier in the semester.

The following table depicts the placement of our samples in the 12-well gel:

Well Sample
1 Ladder
8 30:1 [Au]:[BSA] colloid
9 0.6 g/L lysozyme
10 0.12 g/L lysozyme
11 Unknown B

And the rest is history...

Primary Experiment Dialysis #2 Work-up

30:1 colloid solution vs CaCl2 using MWCO 3500

Well 1 Well 2 Well 3 Well 4 Well 5
30:1 colloid 30:1 colloid 30:1 colloid 30:1 colloid 30:1 colloid
50 mM CaCl2 5 mM CaCl2 0.5 mM CaCl2 50 μM CaCl2 5 μM CaCl2

Necessary Analyses:

  • Bradford - completed, results need to be uploaded
    • 5 colloid samples, 30:1 colloid reference, and Bradford blank
    • 20 μL sample, 200 μL 1:4 Bradford, 780 μL 50 mM Tris/50 mM NaCl

NOTE: There was a bit of a struggle getting a reliable spectrum for the blank. Was run 3 times in order to average/verify. We also made the executive decision to run samples of 50 μL from now on (diluted with 200 μL Bradford and 750 μL Tris/NaCl) in order to increase the precision/accuracy of the results.

  • Fluorescence - not completed
    • 5 colloid samples, 30:1 colloid reference
    • 10 μL of sample in 990 μL of DI water
  • UV-Vis - not completed
    • 5 colloid samples, 30:1 colloid reference
    • 10 μL of sample in 990 μL of DI water
  • CaCl2 ISE - not completed
    • All 10 samples

Primary Experiment Dialysis #3 Set-Up

0.6 g/L Lysozyme vs CaCl2 using MWCO 3500

Well 1 Well 2 Well 3 Well 4 Well 5
0.6 g/L lysozyme 0.6 g/L lysozyme 0.6 g/L lysozyme 0.6 g/L lysozyme 0.6 g/L lysozome
50 mM CaCl2 5 mM CaCl2 0.5 mM CaCl2 50 μM CaCl2 5 μM CaCl2


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