User:Catherine Koenigsknecht/Notebook/Experimental Biological Chemistry/2011/09/21: Difference between revisions

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==Description==
==Description==
#  
# Run 50mL of the supernatent from yesterday over the column at 1mL/min. Collect 10mL fractions.
# Wash the column with 70mL of column buffer at 1mL/min. Collect 10mL fractions.
# Elute the column with 25mL of 10mM maltose in column buffer at 1mL/min. Collect 5mL fractions.
# Run 150mL of the supernatent from yesterday over the column at 1mL/min. Collect 10mL fractions.
# Wash with 75mL of column buffer at 1mL/min. Collect 10mL fractions.
# Elute the column with 75mL of 10mM maltose in column buffer at 1mL/min. Collect 5mL fractions.


==Data==
==Data==
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==Notes==
==Notes==
This area is for any observations or conclusions that you would like to note.
*10mM maltose(100mL) = 0.36g maltose + column buffer
*We did both of our washes until the "Bradford-by-eye" revealed that no more protein was being washed off of the column.
*A "Bradford-by-eye" is mixing 20μL of Bradford dye with 60μL of water and 20μL of a fraction. When the mixture is bright blue protein is present, a lighter blue means there is less protein, and when the mixture is orange little to no protein is present.




Revision as of 10:06, 4 October 2011

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Entry title

Continue to purify using the column.

Objective

Learn how to maintain an OpenWetWare Notebook.

Description

  1. Run 50mL of the supernatent from yesterday over the column at 1mL/min. Collect 10mL fractions.
  2. Wash the column with 70mL of column buffer at 1mL/min. Collect 10mL fractions.
  3. Elute the column with 25mL of 10mM maltose in column buffer at 1mL/min. Collect 5mL fractions.
  4. Run 150mL of the supernatent from yesterday over the column at 1mL/min. Collect 10mL fractions.
  5. Wash with 75mL of column buffer at 1mL/min. Collect 10mL fractions.
  6. Elute the column with 75mL of 10mM maltose in column buffer at 1mL/min. Collect 5mL fractions.

Data

  • Add data and results here...

Notes

  • 10mM maltose(100mL) = 0.36g maltose + column buffer
  • We did both of our washes until the "Bradford-by-eye" revealed that no more protein was being washed off of the column.
  • A "Bradford-by-eye" is mixing 20μL of Bradford dye with 60μL of water and 20μL of a fraction. When the mixture is bright blue protein is present, a lighter blue means there is less protein, and when the mixture is orange little to no protein is present.


Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.



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