User:Anthony Salvagno/Notebook/Research/2009/09/17/Everything Redo (Gel, Extraction, Ligation, Gel, Extraction)

Gel
I am running a gel of the digested pBR322 to separate the two strands. Looks like an incomplete digestion.

Nanodrop Results for Extraction
Long: 34ng/ul --> 21nM Short: 26ng/ul --> 21nM These are in the freezer labeled L and S (for long and short strand respectively). There is another tube labeled S in there that is the same stuff as this S, but one day older (maybe two I can't remember).

No Ligation today
Instead I am going to reanneal T+B, PCR a new anchor, then once I get the BstXI in I will digest the anchor with that and try ligations on Tues.

Anchor PCR
Put 90ul in 5 tubes. Run reaction like on this day.

Re-Annealing
I decided to reanneal T+B just in case that is also bad. The thermocycler is in use so I am going to improvise.
 * 1) I filled a plastic container with water
 * 2) Put it in the microwave and heat until water boils
 * 3) Put eppi in something that can float and place in the hot water
 * 4) Allow water to slowly cool to room temp (just by leaving it on counter top)
 * 5) Done!