Hello all.
Been trying a number of kits and modifications of said kits, to try and extract usable DNA from cyanobacteria (source is HAB lake water). I had "ok" results from a Qiagen - Flexigene kit #51206 but I had to extend incubations and crush the cyanobacteria using a mortar and pestle with liquid nitrogen. I also had to increase proteinase concentrations as well. I ALSO just got done trying a Qiagen Plant-Pro DNeasey kit. With this, I even tried a tube with the small glass beads to hopefully lyse the cells. This produced no DNA.
Before I go to the literature and try a method I wanted to ask if anyone has a good kit or modification of a method from a commercial kit for the extraction of DNA from Cyanobacteria.
THANKS
Andrew
A classical DNA extraction without use of a kit would be best to provide a large quantity of DNA. There are maxi-kits available (whereby you can then concentrate your DNA down), which provide cleaner DNA. These kits are pretty expensive though. Good luck.
Try the Qiagen DNeasy PowerSoil Pro Kit. We use it for cyanobacteria and it works very well. You need to do no modifications and the extraction happens within 3-4 hours max. Also, the DNA can be used immediately for downstream processing.
All the best!
I agree with Prashant Singh. I have been using the same kit with some modifications aiming to increase DNA concentration and purity.
good luck with your extractions!
My suggestion is not to use kit at all if you want more DNA.
Apply the old method of DNA extraction.
This paper is good.
Have look.
https://doi.org/10.1111/jpy.12847
Thank you Mahnaz (and all) for your replies.
I indeed have that manuscript on my desk! I will give it a try if the method I am currently working on does not pan out (from another paper). I know I asked for a kit recomendation, but I am trying some methods I got from some manuscripts first before I pay for the kit. Thanks everyone!
Andrew