Reference no: EM1389255

1. Assume you want to make an agarose gel to run a DNA digestion that you have prepared, but you do not have the reagents needed to make it. First you must prepare a variety of stock solutions that are needed to make 1X TAE.
How would you accomplish the following?
a. Prepare 600 mL of a 40x Tris acetate (MW = 121.14) solution that will have a final concentration of 1mM.
b. Prepare 500mL of a 40x EDTA solution (MW = 372.24) that will have a working solution of 2mM.
c. Now you want to prepare 1L of 20x TAE using your stock solutions from a) and b). What would you do?
d. Now that you've made up your 20x stock solution of TAE using the reagents you just made in questions a. and b., how much of your 20x TAE stock solution and water would you use to make 500mL of 1x TAE?
e. Using your 1x TAE you make two agarose gels, each 40mL. How much of the 1x TAE and agarose would you need
to make a 0.7% gel and a 1.5% gel?
f. You dissolve your agarose into the TAE and allow the mixtures to cool. Calculate how much ethidium bromide
you need to add to the gels. The rate you need to add EtBr is 2µL per 100mL of agarose gel mixture.
How many µL did you add to your gel?
If your EtBr stock is 10mg/mL, how much (in µg) did you add to your 40mL mixtures?