User:Douglas M. Fox/Notebook/AU CHEM-571 F2011 Lab Support/2014/09/30: Difference between revisions
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#Dialysis | #Dialysis | ||
#*Prepare 10 mL 0.5 g/L Lysozyme | #*Prepare 10 mL 0.5 g/L Lysozyme | ||
#*Prepare 2 mL 50 | #*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 | #*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 | #*Prepare 2 mL 50 mM NaCl (if you read ahead, you can use your NaCl calibration stock) | ||
#*Prepare 1 mL 0.25 mM HCl using your 2.5 mM stored stock | #*Prepare 1 mL 0.25 mM HCl using your 2.5 mM stored stock | ||
#*Cut a piece of 3500 MWCO flat dialysis tubing, wet it, and insert it in the 5-well dialysis chamber | #*Cut a piece of 3500 MWCO flat dialysis tubing, wet it, and insert it in the 5-well dialysis chamber | ||
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#**You may need to tip the chamber on an angle to reduce water surface tension | #**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 | #*Fill the other side of the well with one of the following solutions | ||
#**HPLC water, 0.25 mM HCl, 50 | #**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 | #**Each well has a different solution, which will be tested tomorrow | ||
#*Insert top screws to prevent leaking/evaporation | #*Insert top screws to prevent leaking/evaporation | ||
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#*Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM CaCl<sub>2</sub> | #*Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM CaCl<sub>2</sub> | ||
#*Measure Ca<sup>2+</sup> using ISE | #*Measure Ca<sup>2+</sup> using ISE | ||
#*Try measuring 50 μM and | #*Try measuring 50 μM and 50 mM of other salts to check for interferences (Mg<sup>2+</sup> should, Na<sup>+</sup> should not) | ||
#*Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution. | #*Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution. | ||
#Cl<sup>-</sup> calibration curve | #Cl<sup>-</sup> calibration curve | ||
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#*Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM NaCl | #*Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM NaCl | ||
#*Measure Cl<sup>-</sup> using ISE | #*Measure Cl<sup>-</sup> using ISE | ||
#*Try measuring 50 μM and | #*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. | #*Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution. | ||
Revision as of 14:57, 29 September 2014
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TasklistWe will continue the rotation that we started last week.
All groups will set up dialysis.
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