User:Anthony Salvagno/Notebook/Research/2009/08/28/Ligation of pBS to Tether construct

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I am ligating the Sap and Not digested pBS to the tether construct. I don't know if it is clear in the table below, but NpBS is being ligated to the A+A construct and SpBS is being ligated to T+B and anchor (hopefully). I've changed the SpBS protocol. Originally I started with 5nM and added 5nM ever 20 min until 30nM has been attained, but today I just threw in 25nM to see what happens. {{#widget:Google Spreadsheet |key=tRxPpeheSyxNNaJ4FNJLiKQ |width=800 |height=200 }} I am running the ligation for 1.25 hours.

Gel Results

I've used half (10 of 20uL) of each sample to use in a gel. That should equate to about 1ug of DNA (plus weights of anchors and shit) for final concentrations around 25nM. I have half saved in Box#3 as tubes 1 and 2. I need a better label. Dates could be good.

Pbs-ligations-8 28 09.png

So for my best guess, I left the ligations to run a little too long. We had crazy concatamers so I just threw the gel out. Poor wasted DNA.