I have trouble doing the calculations on my study guide, and I need to know how
ID: 9303 • Letter: I
Question
I have trouble doing the calculations on my study guide, and I need to know how to do them for the lab final, can someone help me figure it out? thanks:2. Suppose you did a restriction digest of some DNA, and now you want to run some of your digested DNA on a gel to see if it got cut. Here are the important parameters you used in setting up reaction:
• Initial concentration of DNA sample (before you did the restriction digest): 40 µg/ml
• Amount of DNA sample added to the restriction digest: 10 µl
• Final volume of restriction digest: 40 µl
The uncut DNA is 15kb long. You expect the DNA to be cut at one restriction site, resulting in fragments of 10kb and 5kb.
Now you want to run the gel. You want to load enough DNA on the gel to ensure that you have enough 50ng DNA in the 5kb band. How much of your restriction digest do you need to load on the gel?
Explanation / Answer
You want enough DNA on the electrophoresis gel to see a clear band but not so much that you overload the plate. Usually about 10-20 microliters should suffice.