3-piece ligation: EpBR, BpALS, 5′-bio/int-bio adapter results

Lane assignments:

  1. 1kb ladder
  2. 5′ bio adapter reaction
  3. 5′ bio adapter reaction
  4. int-bio adapter reaction
  5. int-bio adapter reaction

I had to split each reaction into 2 lanes because the volume of the reaction wouldn’t fit in 1 lane.
image
So this reaction failed. I’m guessing the digestion of pALS didn’t work. There is an interesting artifact in this gel though. In lanes 2 and 3 you can see 2 bands between 2 and 3 kb. I have no idea why the DNA would separate like that. There are literally only 3 pieces of DNA in here: a ~30bp piece which wouldn’t be visible in this gel, a 2.5kb piece which is the EpBR fragment and is where the 2 bands are, and the 4kb pALS piece which is above these dual bands.

Oh well, I’ll try again at a future date when I run out of the DNA that I have currently produced.