Week+November+14

__November 14th__ Today we ran another gel of the pGlo double digested in Nhe1 and Sma1. Then we cut the vector and GFP bands out of the gel so that we could purify them using an extraction kit of columns and washes. (The kit dissolves the agarose gel and the columns and centrifuge to catch the pure GFP and vector .) Later, we will do ligation to make sure that our plasmid will relinearize properly.

__November 16th__ Today we nanodropped the vector and GFP purification that Paige and I did on Monday. The nanodrop showed that we had nothing in our purification but contamination. We are unsure what went wrong. When cutting out the bands, we photographed the fragments that we were going to dissolved and were able to see the vector and GFP bands in the gel. Because we had no GFP or vector, we had to start redoing the purification today. Francis purified a set of cultures so that we will have a control. The purification that Francis did came out with the wavelength and concentration that it should. On Monday, Paige and I will purify our other four cultures and hopefully come up with the same results. We also started another digest of our plasmid. Paige and I reworked the volumes of reagents and water to be used so that the double digest works out correctly. While doing the digest, we made more BSA reagent. Later Aprell showed us the spectrophotometer that is used at Notre Dame. In AP Chemistry at St. Joe, we were able to find the frequency of maybe eight samples in an hour; with the machine we operated, we were able to measure the frequency of 96 samples in 10 seconds.

2 microliters Buffer 8 microliters Plasmid 2 microliters BSa 1 microliter Sma1 7 microliters Water (Incubate at 25 degrees F) 2 microliters Buffer 8 microliters Plasmid 2 microliters BSa 1 microliter Nhe1 7 microliters Water (Incubate at 37 degrees F)
 * Sma1 Portion of Double Digest**
 * Nhe1 Portion of Double Digest**