Currently, i got SRL Restriction enzymes for performing a double digestion reaction (NdeI and XhoI). However I doubt on the functionality of the enzyme. I first performed single digestion with my empty vector (pET43a) to check if the enzyme is able to digest my plasmid and incubated for 4 hrs (each enzyme individually). However I did not get satisfactory result. Any suggestion would be helpful.
Did you take your plasmid and run uncut plasmid next do plasmid cut with one enzyme and another lane cut with another enzyme? That is really the way to show if the restriction enzymes are the problem. With a double digest it is hard to know if both digested the plasmid.
Michael J. Benedik Yes I did. I have attached the image. However I am unable to figure it out properly.
Hi there,
There is obviously an issue with the NdeI enzyme from SRL but XhoI looks fine.
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