Part:BBa_K1188000:Experience

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User Reviews

UNIQ8ec91d2898959756-partinfo-00000000-QINU

••••• Immudzen

For the 2013 iGEM competition for the CU-Boulder project we used this part extensively and never had any problems with it. The part has a very strong blue expression and it runs up on a regular 0.5% agarose gel. This made it very useful for protein separation experiments. If you grow it up in a large overnight flask like a 1L container it will look like you don't have anything since it ends up looking like a muddy brown color. Once you spin it down though you should see a strong blue. We tossed out a few of our overnights because we thought they had not grown. This part proved to be extremely useful when debugging digestion, ligation and transformation problems early on. The standard part in the backbones is essentially the same as this but with RFP. But digesting and ligating in this part it was easy to see if we had background carry through since it was red or if we got what we wanted which would be blue. Sometimes we ended up with white colonies and by isolating those we found out that sometimes we were getting double backbones or sometimes more. I would highly recommend this part to anyone in the beginning to make sure your protocols work. Just about every one of our protocols was debugged using this part. http://2013.igem.org/Team:CU-Boulder/Notebook/Protocols We ended up with transformations, digests and ligations that worked nearly 100% of the time by using this part. We also used this part in our protein purification experiments. It turns out that RFP runs down on a gel and AmilCP runs up and they separate very cleanly. We made a freiburg AmilCP part https://parts.igem.org/wiki/index.php?title=Part:BBa_K1188001 so hopefully otherwise will find that useful to fuse to their proteins to color tag them and then use our methods to separate out and purify the protein. This seems like such a simple and obvious part but it sure helped us. We probably transformed it more than any other thing we did. For more information on the characterization of this part and how we purified it see http://2013.igem.org/Team:CU-Boulder/Notebook/Experiments/Experiment_14_1 and http://2013.igem.org/Team:CU-Boulder/Notebook/Protocols/ProteinPurificationAgaroseGel

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UNIQ8ec91d2898959756-partinfo-00000002-QINU