User:Anthony Salvagno/Notebook/Research/2009/11/19/Unzipping construct cleanup

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Well now that I got definitive unzipping construct, I have to run a real gel and gel extract and the Qiaclean it. That will come this afternoon cause I have to go to class right now.

Gel

It is useful to know that I can fit about 55ul in one well with the larger comb that I have, for a 50ml gel. I also noticed a potentially huge problem: IMG 0168.JPG
IMG 0166 1.JPG
The second image is way blurry but it details the problem nicely, while the first image is clear, you can't quite make out what is wrong. The gel is crooked! I have no clue how that happened, but I see it as a problem since my DNA may get pulled out of the gel. I will probably lose up to half my DNA. Well I will have to do this process again anyways.

And in other news I am a disaster today. I may have messed up Brian's PCR reaction, and I forgot to take a picture of the gel before dicing. Oh well. I'm going to have to redo this a ton anyways. Based on yesterday's image I'm guessing I have about 25ng/ul which corresponds to about 9nM. I couldn't really gauge how much I had today because the staining didn't illuminate as much. I think a destain might have been useful afterward but all I care about is seeing the bands enough to cut. Anyways, I'll clean up tomorrow and nanodrop it. Hopefully I get some promising results.

BTW, the Anchor-Anchor construct should have significantly less. I don't think that was as efficient.

Nanodrop Results

I won't be able to do this today, because I screwed up making a taped gel earlier. I also worked with Brian a lot. I will gel purify tomorrow.