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