Hello Everyone,
I'm trying to do three plasmid transformation in my chemically competent cells (NEB turbo cells). Can anyone help me with finding the correct protocol for it? Or if you've done it, can you give me some tips on doing it successfully?
I've been successful in performing double plasmid simultaneous transformation in the same competent cells and the protocol seems to work just fine. I tried to do research on three plasmids at the same time and I can't seem to find any research with this information.
Some general info:
Three plasmids are not huge - generally around 3000bp each.
They are each on different resistance for screening - Amp/Kan/Cm
They each carry separate genes.
The concentration of each plasmid is diluted to 20ng/uL.
Thanks in advance. Really appreciate all of your time.
You probably also have to pay attention to the origin of replication used by the three plasmids.
I only used two plasmids simultaneously before, which in my limited experience, the chance of positive is reduced by hundreds if not a thousand fold. The chance to obtain three in a single round is not impossible, but the chance is not high.
2 maybe 3 highly unlikely to get them in a single transformation
suggesting step by step protocol..
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