Is it possible to use DH5 alpha for in-fusion cloning if the insert (2 kb) and the vector (11.7 kb) are this large?
As far as getting the DH5a to work with the InFusion rxn product, I've tried and had no luck but then I also used them in the earlier tests we did so it could just be inexperience that was the problem. I have had it work well using the included Stellar cells (Clontech) but again this was in later trials. I would recommend making a glycerol stock of the Stellar cells if you get a kit with them included, just in case. I ran out of the chemically competent cells but the electrocompetent ones I made from ones that came in the kit worked fine (I had to EtOH-precipitate or column purify the product).
Hi,
I used HB101 with infusion kit for a similar sized combination of insert and vector and had correct clones for all clones I tested and sequenced. If money isn't a point, just give DH5alpha a try. If money matters, try to get HB101 from a lab near by and use these. If you need a protocol to produce chemically competent HB101 just let me know.
Best,
Kai
Thank you Kai. I shall check with DH5 alpha and the Stellar cells. If it doesn't work will get back to you for the protocol.
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