transfect from full plasmid and transfect from PCR product - (Oct/05/2007 )

Hello everyone


I have a question about transfection. What is the different between transfect from full plasmid and transfect from PCR product any one know and what is the best transfect from plasmid or PCR product?


Thank you very much

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I think transfecting lineal plasmid give you better result that circular plasmid.

But between linear plasmid and PCR product, since they have the complete cassette and are properly purified I think there is no difference.

Any other opinion?

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So you think transfecting linear plasmid give you better result that circular plasmid

From where I digeste the circular plasmid to be linear from which part

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The gene expression levels are almost the same for linear and ccc DNA. PCR products have smaller size and more copy numbers than plasmid per ug DNA. There are several papers on this subject published already.

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the big difference is that the plasmid contains the transcriptional and translational elements for the target gene while the PCR products not.

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you can always linearise the plasmid, and digest out the segment of plasmid you wish to transform. PCR is limited in the lenght of product you can make, so limits the size of the construct you transform. You can build bigger structures in plasmids and make a lot more plasmid then PCR product

Linear DNA does transform better as it has free ends to invade the host dsDNA and trigger homologous recombination. Circular plasmids will float (transiant transformation but if the plasmid has ARS can then survive in the host) and only transform above manner if it has been linearised either by endogenous nucleases or by plasmid breakage during manipulation/ plasmid extraction process.

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