My colleague has come up an idea of using two inserts to ligate into one vector at the same time, which is like : vector-SpeI-Insert A-BamHI-Insert B-NotI-vector
(Spel-Insert A-BamHI, BamHI-Insert B- Notl, Vector digested with Spel and Notl).
Then he transformed the system into E.coli and today the colonies showed up.
Unfortunately, the verification PCR only shows the band of Insert B fragment.
(vector is around 3kbp, Insert B is around 1.8kbp, Insert A is around 700kbp, the verification PCR's predicted size is around 2.5kbp containing the part of Inert A and B) We are confused about the result and I really wish to hear from your suggestions.
if your DNA fragments are PCR products, combine your both fragments (Purified DNA) in a single tube and let them aneal, then insert into vector
Thank you for your suggestion.
My DNA fragments are PCR products which are flanked with different restriction enzyme site. I will try your methods and thank you so much!
- Badges
- Science topic
- Similar topics
- Mathematical Sciences
- Statistics
- Statistical Software
- SPSS