Hi everyone,
I am currently looking at trying to improve a DNA extraction method we use within my lab group. In the lysis stage of the extraction, we add phenol, 20% SDS and sodium phosphate pH 8 to the sample before bead beating. We then spin the sample down and take the supernatant forward for further purification. I am looking into removing the addition of phenol and replacing it with a less toxic alternative so that the method is more portable and applicable to industry.
I was wondering if anyone had any suggestions on what I could replace the phenol with and in relation to this explain how it works in the extraction?
Thanks!
Hello, did you ever get any ideas related to this? Would be interested :)
Hi!
We tried using guanidine thiocyanate, but this lowered the yield quite a lot as it wasn't able to separate the proteins in the milk as well as phenol. Now we have focused on reducing the volume of phenol required and making the method more high-throughput, so ultimately we are sticking with the phenol for this method- just less of it!
Hi Emma,
If you need DNA for ITS sequencing I can show you great protocol for that.
Thanks
Hi Marwan,
Thanks for the suggestion - I haven't done ITS sequencing yet, but I am always open and happy to hear new protocols as they always seem to come in useful!
Hi Emma,
I'd suggest this protocol in case you have large number of samples for ITS as DNA by this protocol as I've tried not really so pure. However, this DNA would be great for tracking multilocus regions such ITS.
GOOD LUCK.
Marwan
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