I am planning on running qPCR but with bacterial DNA. Will the qPCR work with the DNA extracted by boiling culture with DI or lysis buffer? Or I have to extract the genomic DNA using the kit (Purelink)?
Thank you.
You can use DI water and pickup the bacterial colony mix in the water use as template for RT-PCR,
Thanks,
So you think it would be good enough for such a sensitive technique as qPCR? That is what I am planning on doing but I am afraid of any interference that can possibly inhibit the reaction. I know that regular PCR work just fine with this technique but I was not sure about qPCR
Did you consider a simple phenol, phenol chloroform, phenol chloroform isoamyl alcohol extraction, followed by ethanol precipitation? It is an organic extraction which generates organic waste but it would certainly work well.
Hi Thanh!
I usually use a kit (AxyPrep) to extract DNA from plant bacteras for qPCR applications, but some times I use CTAB protocol without phenol and it works too. I compared the sensibility of the essay using DNA extracted by both methods but with the same sample and I found small differences in the Ct values.
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