How much of plasmid should be cut to see the gene - (Jan/04/2012 )
Hello, i would like to know how much of the plasmid (Carrying my Gene) should be cut to see my gene on the gel? I have a plasmid of 5.6KB and my gene is o.66kb. In short i am asking what is the minumum amount of the plasmid should i cut so that i can see my gene on the gel?
I guess it depends on the sensitivity of the DNA binding dye used in your gel. I use ethidium bromide, and find it hard to visualize a band less than 20ng. So you would have to digest atleast 170ng of plasmid DNA to get 20ng of insert.
Keep in mind this is the minimum amount you need though..if you have enough of your plasmid, just simply digest 500 or 1000ng of your plasmid and run it in one go to see a nice insert band.