KRC Reagent Calculator

	A	B	C	D	E
1	KRC Reagent Calculator
2
3	1. Enter final volume:	500	ml
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5	2. Start with this amount of purified water:	350	ml
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7	3. Add these reagents:	Amount (g)		FW (g/mol)	[final] (mM)
8	NaCl	3.448		58.44	118
9	KCl	0.175		74.55	4.7
10	CaCl2·2H2O	0.147		147.02	2
11	MgSO4·7H2O	0.493		246.48	4
12	D-glucose	0.991		180.16	11
13	NaHCO3	0.525		84.01	12.5
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15	5. Add this amount of purified water into a separate beaker:	25	ml
16	And add this amount of Na2CO3:	0.6625	g	106	12.5
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18	6. Add Na2CO3 solution slowly (1-2 ml at a time), while adjusting the pH between 7.5 and 8 with 1N HCl.
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20	7. Allow the solution to mix for several minutes before adjusting the pH to 7.35-7.40, then bring to the final volume with purified water.
21
22	8. Filter through a vacuum filtration system and place in a 1- or 2-L bottle.
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24	This calculator works for a perfusion-fixation protocol developed by the Kristen Harris Lab at The University of Texas at Austin.
25	See the full protocol here:	https://wikis.utexas.edu/display/khlab/Transcardiac+Perfusion
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