Calculate BOTH working concentrations of DMCU. I was given this formula to use:

Question

Calculate BOTH working concentrations of DMCU.

I was given this formula to use: C1V1=C2V2

Where C1 is the original or stock concentration, V1 is the volume of the stock added, C2 is the final or working concentration and V2 is the final volume.

And this information:

a. 2x + 7 x 30. S. * 31. - V + ? de 4 PART 2: INHIBITION OF HILL REACTION BY HERBICIDE DCMU Note: For Part 2, you will test whether the pesticide DCMU interferes with electron transport during the Hill Reaction. You will NOT do this part in triplicate. 1. Obtain 3 clean test tubes. 2. Place all 3 tubes on ice and label one each "C" "low," and "high." Add 3.5 ml di- luted chloroplasts to each. Lay a piece of foil over the top of the ice bucket. 3. Make a dilution of DCMU from the DCMU stock solution as follows (DCMU stock is 0.1 mM): a. Label a clean microfuge tube “10 M DCMU." b. Using a P200, add 100 pul H,O to the microfuge tube. C. Using a P20, add 11 ul stock DCMU to the microfuge tube. Close cap and invert to mix. 4. You are now ready to add DCMU to the reaction tubes on ice. Leave all tubes on ice after addition. For this experiment, your control is a tube in which you allow photosynthesis to proceed normally. Add 100 pul H,0 to the “control" test tube on ice. H,O should not affect photosynthesis and is used to ensure that the reaction volume is the same in all tubes. 5. Add 100 uL 10 puM DCMU to the "low" test tube and mix. 6. Add 100 mL stock DCMU (0.1 mM) to the "high" test tube and mix. 7. Re-cover all tubes with foil and wait 10 minutes to allow the DCMU to enter the chloroplasts and bind to Photosystem II. 8. Add 500 uL DCPIP (0.4 mm) to each tube and mix. 9. Remove foil and measure the A600 of each of the three samples and enter into your lab notebook. These will be your Time O readings. 10. Assay the Hill Reaction every 2 minutes as you did in Part 1. Keep all tubes under fluorescent light when not reading absorbance. Enter all data into the appropriate table in your lab notebook. CLEAN-UP 0 Remove test tubes and turn off Spec 20. O Pour all DCPIP waste into DCPIP waste container. O Rinse all test tubes and dispose of rinsate in DCPIP waste container. O Put all used serological pipets, tips down, into pipet container. o Throw away pipet tips into the garbage. O Submerge test tubes under water in wash pan. 1 1' u oftono in trash and submerge tubes in dishpa

Explanation / Answer

The working concentration of DCMU is 10 micromolar for LOW tube.

And the working concentration of DCMU is 0.1 mM for HIGH tube.

The 10 micromolar is made from 0.1 mM DCMU using the equation C1V1=C2V2

To make 100 microlitre 10 micromolar DCMU -

C1 = concentration of DCMU - 10 micromolar

V1 = volume of DCMU - 100 microlitre

C2 = concentration of stock DCMU - 0.1 mM = 0.1 * 1000 micromolar = 100 micromolar

C1V1=C2V2

C2V2=C1V1

V2=C1V1/C2

=(10*100)/100

=10 microlitre

So from stock 0.1 mM DCMU you have to take 10 microlitre and pipet it out in a 100 microlitre tube and fill the rest (90 microlitre) with water.

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