Question

Effect of varying enzyme concentration

1. Prepare a series of diluted enzyme concentration in 0.5 mL eppendorf tubes. Use the DEA buffer as the diluent. Use enzyme concentrations of about 1/5 to approx. 1/35 of the stock enzyme. One suggestion is to serial dilute 20 micro liters of a 1/4 dilution by 1/2 to give 20 micro liters each of 1/4, 1/8/ 1/16, 1/32 dilutions. How would I go about calculating and showing this and if possible show step by step guides.

Answer 1

Given: Serial dilute 20 µl of a 1/4 dilution by 1/2 to give 20 µl each of 1/4, 1/8/ 1/16, 1/32 dilutions.

Stock solution is 20 µl. The stock is already the ¼ dilution.

Now, take 10 µl of this stock solution and mix with 10 µl of distilled water. So, the new concentration becomes half of the initial ¼^th dilution = ½ * ¼ = 1/8.

Now, take 10 µl of this 1/8^th dilution and mix with 10 µl of distilled water. So, the new concentration becomes half of the initial 1/8^th dilution = ½ * 1/8 = 1/16.

Now, take 10 µl of this 1/16^th dilution and mix with 10 µl of distilled water. So, the new concentration becomes half of the initial 1/16^th dilution = ½ * 1/16 = 1/32.