Question by Ron W: What methods can be used to verify that transformation occurred?
Besides using ampicillin resistance, what methods can be used to verify that transformation occurred?
Best answer:
Answer by kwicher
If you want to know in general what selection method one can use:
– resistance to other antibiotics (chloramphenicol, kanamycin, zeocin )
– use of auxotrophic strains, which are unable to grow in the absence of some particular compound and plasmids which provide gene allowing for synthesis of this compound
(http://en.wikipedia.org/wiki/Auxotrophic)
Give your answer to this question below!
Well, transformation requires the transfer of something. So far you have talked about the thing you can see, amp resistance. That would be the ability to grow on a plate with amp. but what is being transfered? If it is a plasmid, then you could look for the plasmid (plasmid mini prep, digest, run the gel). If it is some other genetic material you could look for the presence of that genetic material (genomic DNA prep, digest, southern transfer and probe for the new stuff). You don’t give enough info here to know what is being transformed, but in the case of a plasmid, amp resistance is not enough. You can from time to time find spontaneous mutants that will grow on amp (not many, but it’s happened to me). In those cases they can grow a little on the plates, but don’t do well at all in liquid.