Date: Mon, Oct 26, 2009 at 3:30 PM
Now we get to the sticky part of the class. Our experiments should work, yet in the past I've had poor overall success. What we will be doing is indeed trial and error, such is science. On Tuesday I will lecture a little on DNA and then we'll set up restriction digestions of our genomic DNA. There is no quiz.
So I need you to calculate the following:
1. How much genomic is needed to put 2ug in a tube to reach a FV of 15ul with the addition of water
2. How to set up a restriction digestion given the following information:
2 ug of genomic DNA
1ul of EcoRI
Xul of 10x buffer
FV = 15 ul
3. Set up a SalI digestion:
10ug of genomic DNA
15u/ug of enzyme to DNA if the enzyme comes at a concentration of 10u/ul
how much 10x buffer?
how much water?
reach some final volume (it should be around 150ul)
So in other words, review how to set up digestions and we'll do it at the end of class. I've mapped out a strategy for the next 3 classes, cross your fingers!