What strain of e.coli are you transforming? The usual cloning host like e. Coli DH10b, dh5alpha don't have their own plasmids. So, there is no such issue.
Ok Thank You Sir! actually i am using E. coli DE3 commercial strain ,.... i think that will be the reason , Welcome.
Generally not recommended to isolate the plasmid from expression hosts, it may contain plasmid for special codons or expression regulation, chaperones etc. In that case, you will get both the plasmid and there is no way to separate them during isolation. One way I can think of is to digest the isolated plasmids with a single restriction enzyme known for sure to cut at a single site of the target plasmid, which would linearize the vector so that it can be identified on the gel against the DNA ladder and gel purified. Ligate and retransform.
If the plasmid position is known for sure than restriction digestion step not required and the target band can be gel purified.
Mangal Singh ! Ok all right but i am confused in one thing is that! once when we have integrated our GOI into the plants genome by using Agrobacterium! How we can make the cDNA from the gene once integrated into the genome of the plants cells? for QPCR or Semi quantitative PCR? please tanx!
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