User:Michael S. Bible/Notebook/571/2014/09/30: Difference between revisions

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==Objective==
==Objective==
Learn how to maintain an OpenWetWare Notebook.
Created a create a Ca<sup>2+</sup> calibration curve and began a new dialysis


==Description==
==Description==
# Add experimental record here. Include what, how, and why...
# CaCl<sub>2</sub> Calibration Curve
#*Prepare 2 mL 50 mM CaCl<sub>2</sub> (if you read ahead, you can use your CaCl<sub>2</sub> calibration stock)
#*Prepare 2 mL 500 μM CaCl<sub>2</sub> (if you read ahead, you can use a CaCl<sub>2</sub> calibration dilution)
#*Prepare 2 mL 50 mM NaCl (if you read ahead, you can use your NaCl calibration stock)
# Cl<sup>-</sup> calibration curve
#*Prepare 25 mL 50 mM NaCl using HPLC water
#*Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM NaCl
#*Measure Cl<sup>-</sup> using ISE
#*Try measuring 50 μM and 50 mM of other salts to check for interferences & ion-pairing
#*Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution.
# Dialysis
#*Prepare 10 mL 0.5 g/L Lysozyme
#*Cut a piece of 3500 MWCO flat dialysis tubing, wet it, and insert it in the 5-well dialysis chamber
#*Clamp the dialysis chamber shut
#*Fill one side of each well with 1 mL of your 0.5 g/L Lysozyme solution
#**Transfer 1 mL to a plastic test tube using a pippetter, then
#**Carefully fill each well with with a glass Pasteur pipette
#**You may need to tip the chamber on an angle to reduce water surface tension
#*Fill the other side of the well with one of the following solutions
#**HPLC water, 0.25 mM HCl, 50 mM CaCl<sub>2</sub>, 500 μM CaCl<sub>2</sub>, or 50 mM NaCl
#**Each well has a different solution, which will be tested tomorrow
#*Insert top screws to prevent leaking/evaporation
 
 


==Data==
==Data==
* Add data and results here...
* For the 0.5 g/L Lysozyme, 0.00570g Lysozyme used.
* For NaCl serial dilution 0.0713g NaCl was placed in 25 mL of water
* For the 0.25 mM HCl, a new solution was made containing HPLC water rather than DI water


==Notes==
==Notes==

Revision as of 15:13, 1 October 2014

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Objective

Created a create a Ca2+ calibration curve and began a new dialysis

Description

  1. CaCl2 Calibration Curve
    • Prepare 2 mL 50 mM CaCl2 (if you read ahead, you can use your CaCl2 calibration stock)
    • Prepare 2 mL 500 μM CaCl2 (if you read ahead, you can use a CaCl2 calibration dilution)
    • Prepare 2 mL 50 mM NaCl (if you read ahead, you can use your NaCl calibration stock)
  2. Cl- calibration curve
    • Prepare 25 mL 50 mM NaCl using HPLC water
    • Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM NaCl
    • Measure Cl- using ISE
    • Try measuring 50 μM and 50 mM of other salts to check for interferences & ion-pairing
    • Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution.
  3. Dialysis
    • Prepare 10 mL 0.5 g/L Lysozyme
    • Cut a piece of 3500 MWCO flat dialysis tubing, wet it, and insert it in the 5-well dialysis chamber
    • Clamp the dialysis chamber shut
    • Fill one side of each well with 1 mL of your 0.5 g/L Lysozyme solution
      • Transfer 1 mL to a plastic test tube using a pippetter, then
      • Carefully fill each well with with a glass Pasteur pipette
      • You may need to tip the chamber on an angle to reduce water surface tension
    • Fill the other side of the well with one of the following solutions
      • HPLC water, 0.25 mM HCl, 50 mM CaCl2, 500 μM CaCl2, or 50 mM NaCl
      • Each well has a different solution, which will be tested tomorrow
    • Insert top screws to prevent leaking/evaporation


Data

  • For the 0.5 g/L Lysozyme, 0.00570g Lysozyme used.
  • For NaCl serial dilution 0.0713g NaCl was placed in 25 mL of water
  • For the 0.25 mM HCl, a new solution was made containing HPLC water rather than DI water

Notes

This area is for any observations or conclusions that you would like to note.


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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