So I have a gene of interest inserted into the pET28a plasmid and I've used it in colony PCR with BL21 cells. The primers I used were pET28 primers. After running the agarose gel with my samples I saw a band similar to the size of my insert. Can someone explain this to me...I thought I would see a band that's the size of my plasmid+insert.
when you say that you are using pET28 primers, that is not enough information to troubleshoot your experiment. If the primers on opposite sides of the insert you would only amplify the insert (assuming the primers are oriented towards each other and flanking the insert). However if the primers are adjacent to each other and not flanking the insert then you might expect to amplify the plasmid plus insert.
More details are needed. What is the size of your insert and where are the primers in relation to it. For example if your insert is 2kb and the primers are 20 bases either side of it then you will not see the difference in size unless the gel runs a very long way. You will only see whole plasmid plus insert by single restriction cutting the inserted plasmid and running that.
You need the map of vecor, set primer on it and calculate the size of recombinant vector and compare with nonrecombinant vector.
You could find the vector map on addgene or snapgene easily.
Without map, more detail needed about place of primers.
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