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Changing the vector

cloning

30 replies to this topic

#31 Bio-Lad

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Posted 28 August 2014 - 06:54 PM

After ligation you have to transform to your competent cells and plate them to selective plates to kill of those without plasmid. Then you generally culture a few of the colonies on your plate in liquid medium, do a miniprep to extract the plasmid (keep some of the culture to make a frozen glycerol stock if the clone turns out to be good) and cut with a restriction enzyme that will give different patterns with and without the gene of interest. Send the clones that look correct for sequencing.

Try pLOT, the a free plasmid mapping tool!

http://plasmidplotter.blogspot.com/






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