Hi. I want to ligate a small DNA fragment (a luminescence tag), 33bp into a vector 3.3kbp.
I do not know how. So I am wondering if I can design just one primer that covers that whole DNA fragment with RE sites included. Then I run PCR with sample (another vector that contains the tag). So double stranded DNA of the 33bp fragment should be produced. Then I will digest the purified PCR product with RE.
But instead of run gel and excise the band (because I do not think I can detect such small size of DNA in gel), can I just directly use the RE digested mixture and ligate to the linearized vector? And expect the 33bp fragment can ligate with the vector?
Edited by Thomson, 13 March 2018 - 11:20 PM.