Hi,
Cryptosporidium has robust and hardy cell wall and I wonder if any commercial DNA extraction kits are able to extract its DNA. Some studies conducted freeze-thaw step in liquid nitrogen before proceed with DNA extraction utilising a DNA extraction kit.
Whereas, is it necessary because some other studies did not employ freeze-thaw method. Or is it some kits are 'superior' and do not need additional freeze-thaw step beforehand?
Thanks
Hello Syatir, I've never worked on the extraction of Crytosporidium, however most of the DNA extraction protocols that I've read for the Cryptosporidium oocyst use the freeze-thaw. However, you can read this paper below where they used direct extraction of the DNA and compare to those methods using freeze-thaw. Good luck.
http://aem.asm.org/content/71/3/1135.full
Hi Syatir, May I ask you what is ur purpose to DNA extract oocysts? Just only oocysts or oocyst in mice feces? Why dont you count the number of oocsyts then make excystation to get sporozoites, then DNA extraction is easier. With mice feces We use ultrasonication first then use Qiagen kit for futher step.
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