User:Anthony Salvagno/Notebook/Research/2010/06/25/Unzipping Stuff:Gel Extraction and Gel analysis

Quick Navigation: Pages by Category | List of All Pages | Protocols

I tried this stuff yesterday (I'm predating this so technically it is today) and it all failed. I did a massive digestion and ruined like half my batch by gel extracting the wrong piece. I gel extracted the right piece later. I also tried the nuetravidin-adapter bonding and I didn't add enough Neutravidin, another fail. I'll make amends and do it all right!

Biotin-Avidin Bonding

See these notes. 1mg/ml is my stock of Neutravidin and I am starting with 10um .025 dsDNA with overhangs. According to my notes the stock is ~16uM so I can do a 1:1 reaction and then run a gel.

Gel of pBR Digest

I also gel extracted the wrong piece piece of DNA so I will have to run a diagnostic gel of the gel extracted stuff to make sure I'm right. I will then simultaneously gel extract the piece that I messed up in case I am right. Fun in the sun.

Ligation Reaction

I am determined to finish the ligations today (tomorrow) so once the diagnostic checks out I will be doing this reaction. If it fails then it will have to wait.