Question

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Final             Volume of              Volume of               Volume of       Final volume (uL)

[PNPP] mM   0.5mM PNPP(ul)   0.2M Tris-HCl(ul)   enzyme (uL)

0.01                                                                            100                        1500

0.02                                                                             100                       1500

0.04                                                                             100                         1500

0.06                                                                            100                            1500

0.08                                                                            100                              1500

0.1                                                                           100                             1500

0.2                                                                              100                               1500

0.3                                                                           100                               1500

0.4                                                                             100                                1500

Answer 1

1. total volume of solution = 1500micro L

Volume of enzyme = 100 micro L

Volume of 0.5mM = 0.01 X 1500 / 0.5 = 30microL

so volume of 0.2 M Tris HCl = 1500-(30+100) = 1370 micro L

2. total volume of solution = 1500micro L

Volume of enzyme = 100 micro L

Volume of 0.5mM = 0.02 X 1500 / 0.5 = 60microL

so volume of 0.2 M Tris HCl = 1500-(60+100) = 1340 micro L

3.total volume of solution = 1500micro L

Volume of enzyme = 100 micro L

Volume of 0.5mM = 0.04 X 1500 / 0.5 = 120microL

so volume of 0.2 M Tris HCl = 1500-(120+100) = 1280 micro L

4. total volume of solution = 1500micro L

Volume of enzyme = 100 micro L

Volume of 0.5mM = 0.06 X 1500 / 0.5 = 180microL

so volume of 0.2 M Tris HCl = 1500-(180+100) = 1220 micro L

5. total volume of solution = 1500micro L

Volume of enzyme = 100 micro L

Volume of 0.5mM = 0.08 X 1500 / 0.5 = 240microL

so volume of 0.2 M Tris HCl = 1500-(240+100) = 1160 micro L

6.total volume of solution = 1500micro L

Volume of enzyme = 100 micro L

Volume of 0.5mM = 0.1 X 1500 / 0.5 = 300microL

so volume of 0.2 M Tris HCl = 1500-(300+100) = 1100 micro L

7. total volume of solution = 1500micro L

Volume of enzyme = 100 micro L

Volume of 0.5mM = 0.2 X 1500 / 0.5 = 600microL

so volume of 0.2 M Tris HCl = 1500-(600+100) = 800 micro L

8. total volume of solution = 1500micro L

Volume of enzyme = 100 micro L

Volume of 0.5mM = 0.3 X 1500 / 0.5 = 900microL

so volume of 0.2 M Tris HCl = 1500-(900+100) = 500 micro L

9. total volume of solution = 1500micro L

Volume of enzyme = 100 micro L

Volume of 0.5mM = 0.4 X 1500 / 0.5 = 1200microL

so volume of 0.2 M Tris HCl = 1500-(1200+100) = 200 micro L