CTAB DNA extraction for Salmonella - (Mar/27/2007 )

Hi all,

I have been trying to isolate DNA from Salmonella using CTAB method. I've been using the CTAB method for plants and it works nicely. The same CTAB method also works for Bacillus, but I got almost no DNA with Salmonella. Freeze thawing three times helps but it only increase the amount of DNA by a tiny amount. The DNA would eventually be used as PCR template, and I think tiny is enough but if I were to do a digest and run DNA electrophoresis on it, it won't be give me clear bands. I am aware of the different DNA extraction methods that I can try, but since the lab is new, and it takes 1-2 months to order anything in this part of the world, I gotta find a way to make this method works in the mean time.

Any suggestion?

If you are using this as a PCR template, you are doing far too much work. Simply add a very small amount of cells. Use a pipet tip touched to a colony vortexed into 50 ul of water, and add 1 ul to the PCR reaction. Extend the initial 95C part of the PCR cycle to 5 minutes and go.

You won't ever get clear bands from digesting genomic DNA except in rare cases.

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hello there is one modified CTAB method you can try that
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to coffee:
hello, would u kind enough to tell me that modified CTAB method.My experiment now was in urgent phase. Thanks very much.