User:Michael F. Nagle/Notebook/Chem 571/2013/01/30

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(Preparation of ADA aliquots)
(Preparation of phosphate buffer)
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==Preparation of phosphate buffer==
==Preparation of phosphate buffer==
*3.72g NaH2PO4 and 13.08 Na2HPO4 were added to 1.2L water, which was shaken.
*3.72g NaH2PO4 and 13.08 Na2HPO4 were added to 1.2L water, which was shaken.
-
*pH was brought to 7.4 NaOH
+
*pH was brought to 7.4 using NaOH
==Preparation of ADA aliquots==
==Preparation of ADA aliquots==

Revision as of 14:58, 30 January 2013

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Objective

  • Prepare stock solutions of adenosine and phosphate buffer
  • Prepare aliquots of ADA stock solution
    • Stocks will be used for ADA activity assay next week

Preparation of adenosine stock solution

  • 10mL of 15mM solution of adenosine was prepared
    • .015M = x/.01
    • 0.0015mol adenosine * 267.24mol/g adenosine = 401mg adenosine
    • 401mg adenosine was weighed and added to 10mL phosphate buffer.
  • The 15mM solution was diluted to 5mM
    • .01L * .015M = .005M * x
    • 0.03L total volume for 5mM
    • 0.02L buffer was added to bring the solution to 30mL and 5mM

Preparation of phosphate buffer

  • 3.72g NaH2PO4 and 13.08 Na2HPO4 were added to 1.2L water, which was shaken.
  • pH was brought to 7.4 using NaOH

Preparation of ADA aliquots

  • Dhea Patel measured out 1.7g 0.5unit/mL ADA
  • Stock solution was prepared by adding 68 mL phosphate buffer to the ADA
  • This was divided into 68 1mL centrifuge tubes, which were stored in the freezer.




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