BL21 Plasmid extraction? - (Jun/06/2008 )
Can you extract Plasmid from BL21 using a miniprep kit? If you can how pure do you think it is? Thanks
See these links in case you find an answer, no personal experience!:
Extracting plasmid DNA from BL21 cells is a bad idea. This strain is endA+, and expresses an endonuclease which damages DNA. You would be much better off extracting plasmid DNA from a cloning strain such as DH5a, DH10B or other endA- strains. If you do extract from BL21, I would make certain to do the extra washes in the miniprep protocols, and, if I wanted the DNA for long term storage, I would retransform into a cloning strain and miniprep from that.
well we have this dna made for us which uses the plasmid pet dest42 which cant use dh5a cell for some ccdb effects. This is ruining my world now, and my boss just had his grant expire and i didnt catch this before the grant expired which means no cash for the 200 dollar cells required to clone them. Does anyone have any better ideas to efficiently clone the dna? I mean i could always try and put into a new vector but that costs money as well and time to learn how to do since i havent. Thanks
hello thanks very much for your wonderful explaination, but I have one question: if the strain is endA+ which damage DNA, how can it translate to protein, for it has no DNA then?