User:Douglas M. Fox/Notebook/AU CHEM-571 F2011 Lab Support/2014/09/30: Difference between revisions
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== | ==Tasklist== | ||
* Insert | We will continue the rotation that we started last week. | ||
All groups will set up dialysis. <br> | |||
1 Group will work on GPC. <br> | |||
The other two will either retry Bradford, create a Ca<sup>2+</sup> calibration, or a Cl<sup>-</sup> calibration. | |||
<br> | |||
<br> | |||
#Dialysis | |||
#*Prepare 10 mL 0.5 g/L Lysozyme | |||
#*Prepare 2 mL 50 μM CaCl<sup>2</sup> (if you read ahead, you can use your CaCl<sup>2</sup> calibration stock) | |||
#*Prepare 2 mL 50 μM NaCl (if you read ahead, you can use your NaCl calibration stock) | |||
#*Prepare 2 mL 50 μM KCl | |||
#*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 | |||
#*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 μM CaCl<sup>2</sup>, 50 μM NaCl, or 50 μM KCl | |||
#**Each well has a different solution, which will be tested tomorrow | |||
#*Insert top screws to prevent leaking/evaporation | |||
#GPC | |||
#*Prepare 500 mL 0.5M HAc / 0.3M Na<sub>2</sub>SO<sub>4</sub> | |||
#*Prepare 10 mL 1 g/L Lysozyme using the HAc/Na<sub>2</sub>SO<sub>4</sub> stock | |||
#*Using your 1 g/L Lysozyme & HAc/Na<sub>2</sub>SO<sub>4</sub> stock, prepare 5 mL 0.5 g/L & 5 mL 0.1 g/L Lysozyme | |||
#*Filter your remaining HAc/Na<sub>2</sub>SO<sub>4</sub> stock through 0.2 μM membrane filter into clean & dry filter flask | |||
#*Run GPC (I will instruct you) | |||
#Ca<sup>2+</sup> calibration curve | |||
#*Prepare 25 mL 50 mM CaCl<sub>2</sub> using HPLC water | |||
#*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 | |||
#*Try measuring 50 μM and 5 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. | |||
#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 5 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. | |||
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TasklistWe will continue the rotation that we started last week.
All groups will set up dialysis.
|