From 2012.igem.org
Team:Technion/25_June_2012
Ilya
Today was one of these days everything goes wrong. We had two reactions to cut the pLux promoter. We intended to use one as it is and the gel purify the other. When I tried to load the first sample to the gel, I mixed it with loading buffer on the parafilm (which was a mistake since I had 15 ul. we are supposed to mix only 5 ul on parafilm, not more). I managed to fill the sample with bubbles and wasn't able to load it. Eventually I lost the sample. So I tried again, this time mixing the loading buffer in the ependorf and with some help, I managed to load it. However, when I looked at the gel in the UV camera, I didn't see the expected band at 55bp at all. I think that it's concentration was too low. We used 1 ug for the restriction but the insert was only ~1/40 from the size of the complete plasmid, meaning we had 25ng of the insert. No wonder we didn't see it in the UV camera, it's not enough DNA to see.
Since we lost all of the source for our insert, I put starters from the glycerol stock of the bacteria with the plasmid. I set up two starters of 5 ml each for miniprep tomorrow.
I also cleaned the pSB1C3 backbone which we cut yesterday. We started with 8 ul of 25 ng/ul stock which was supplied by the registry. This means we had 200 ng of linear plasmid. After cleaning I got the following results:
20 ul of 11.8 ng/ul and 10 ul of 6 ng/ul DNA. This is very weird because we get over 200 ng of DNA. Something went wrong. Either the concentration of the stock was wrong and we began with more than 200 ng or something went wrong during the cleaning process. I am not sure whether or not these samples are going to be useful or not. We will ask Sharon what to do later.
One thing that I actually managed to do today without messing it up was editing the wiki a little. I managed to set up our notebook with a calendar. I also created a template for the navigation bar and another template for the notebook. We will update these templates later to give our pages a better look. For now, I am aiming for functionality and not styling.
Inbal
Asaf
Hila
Lior
Noa
Evgeni
Shahar
Rachel
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