User:Anthony Salvagno/Notebook/Research/2009/09/30/Brian's Ligation Results
User:Brian P. Josey/Notebook/2009/09/30 contains the results of the gel from the ligation. Also I have some notes on there about what we are seeing.
I cut the slice (upper band) from this gel and I think I may have gotten some of the lower band (1.8kb) in my slice. Although now I am skeptical because that band looks a lot like 1.5kb not like 1.8kb. Maybe there could also be some kind of weird concatamers with the adapter duplex, but I find that unlikely.
In any event the ligation failed and I think there are 2 possible problems:
- no BstXI digestion of anchor
- no T+B annealing
Maybe the T4 Ligase is choking the chicken as well, but who knows. Koch, any thoughts?
- Steve Koch 02:49, 1 October 2009 (EDT): After just reading your other comments, I came to same conclusion that it's just leftover from the gel extraction. I do have thoughts, and we should talk about them tomorrow. Basically just systematically approaching this problem. I know you can get it soon. One way of looking at my thoughts is I want to do more planning and less reacting to results. A trap that I commonly fall into.