Week+of+October+10

__October 10th__ Today, we used the minipreps that we made last Wednesday and did a nanodrop to test the DNA concentration. We found the concentration was 70-80%. Then, we used the green mix and Aprell's taq to do another PCR, using the same procedure as before (Procedure). The results will be ready for Wednesday.

__October 12th__ Using the PCR products from the PCR of the mini preps from Monday, we ran another gel and found that the first colony worked well but the second colony did not work as well.

(photo from google images ) ^ (picture of what it looks like to run gel)

We then used these colonies to do a digest in which we will use restriciton enzymes to cut the GFP out of the plasmid in order to use and reagents to use in PCR again. (That is, we are running the lab in reverse. We are starting with the plasmid and working backwards to get just the linear GFP ) The two restriction enzymes that we are using are Eco R1 and Age1. These enymes will read the plasmid and only cut in the specific sites for which they are coded.

For the digest, we added the following to our Plasmid. The solution will be left overnight to digest in order to be sure that we will have almost pure GFP left with which to do PCR.
 * 1) 2 milliliters of Buffer
 * 2) 5 milliliters of Plasmid
 * 3) 1 milliliter of Age1
 * 4) 1 milliliter of EcoR1
 * 5) 2 milliliters of BSA
 * 6) 9 milliliters of Water