User:Anthony Salvagno/Notebook/Research/2009/05/07/Successful Ligation!

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I've done it! I ran a super quick gel (on a glass plate) and loaded the gel with 2uL of ligation product (from Tues). It can clearly (by clearly I mean poorly) be seen that one of the samples has a successful ligation. For those that have no idea what that image is let me break it down:

  1. The first lane is the 1kb ladder.
  2. Next is ligation of anchor + SapI adapter + Sap14 (my naming convention for pBS with random fragment from tube 14 digested with SapI).
  3. Next is ligation of anchor + SapI adapter + Sap16
  4. Ligation of anchor + SapI adapter + Sap21
  5. Ligation of anchor + SapI adapter + Saplib

We did not use SapD because there were several SapI sites and we wanted to get the ligation right before we did tougher ligations.

By the way, The Sap16 lane is the one that is the obvious success. There are 3 bands because the bottom band is just anchor + adapter. The next one up is just plasmid, and the top is the ligation event! Awesome. Kelly is destaining and will take another picture to see if the poor image is because of lack of DNA or because of lack of proper staining. The Saplib lane shouldn't make sense since it is a conglomeration of many random fragments.

  • Ramalldf 04:43, 13 May 2009 (EDT):WOW. Congratulations, this is huge. I don't know how ready the trap is yet, but this result basically means that shotgun unzipping is pretty much ready, no?
  • Anthony Salvagno 12:44, 13 May 2009 (EDT): Yea the trap is days away from being completed (minus calibrations and stuff). There is no stopping us now. Wish you could be here for this.