In-Fusion

[Iona Christie]

Has anyone else used the TaKara In-Fusion kit? I have tried it for the first time and have gotten no colonies on my plates; the control has worked. I am using  One Shot™ Stbl3™ Chemically Competent E. coli. Any thoughts on why it has not worked are welcome.

[servit...@gmail.com]

I'm actually in the process of using it right now. Our lab recently returned to a project that we first started in 2019, in which we're trying to use In-Fusion to assemble four PCR products into Takara's Adeno-X vector. Our success rate with In-Fusion has been somewhat hit-and-miss, although to date I'm not sure how much of that has been issues with In-Fusion itself and how much has been other factors.

Can you tell us more about your experiment? What vector are you using? How many PCR inserts? What polymerase are you using to create your inserts? Have you used Takara's In-Fusion Molar Ratio Calculator to ensure you're combining your vector and insert(s) at the correct ratio?