User:Anthony Salvagno/Notebook/Research/2009/04/24/Sequencing and Transformation of inserts

From OpenWetWare

Jump to: navigation, search

I kind of left this day blank because I didn't do much, but I feel like I should write something.

Sequencing

We are doing two plans of attack right now:

  1. Digest the plasmid with NotI and ligate the fragment to an anchor hairpin via another NotI digestion. So basically, anchor(BstXI)adapter(NotI)fragment. I will make pictures later.
  2. Digest the plasmid with SapI and ligate to anchor and annealed oligos with SapI overhang. anchor(BstXI)adapter(SapI)fragment.

We sent the DNA off to get sequenced and postponed the SapI digestion because of low DNA.

Transformations

I've been using letter and numbers to label my DNA. Letters represent genomic DNA digestion with XhoI and then ligated to XhoI digestion pBS and the transformed. Numbers represent gDNA digested with XhoI+EcoRI and then ligated to double digested pBS and then transformed.

I ran low on the DNA that was used for the NotI test (we tested to make sure there were no cut sites within the framents) and those tubes were D, 2, 16, and 21. I also want to digest tubes D, 14, 16, and 21 with SapI. We sent tubes D, 2, 14, 16, 21 for sequencing. I did a transformation for those 5 tubes.

Personal tools