Large plasmid ligation: do i need Alkaline phosphatase?
Hi everyone, i'm a new user.
I have a doubt about a protocol I'm using for a ligation of a very large plasmid (14kbp) with an insert: after digestion with a single-cut enzyme (sticky ends), the protocol passes directly to the purification of the plasmid without making any alkaline phosphatase. My doubt is during the ligation, the plasmid will self-ligated without accepting the insert . Am I mistaken or the protocol is incorrect?