We shall collect biopsies from skin/hoof of animals for bacteriological microbiome analysis. We need suggestions on DNA extraction kit that may be used for this. We prefer to keep the biopsies in a DNA stabilizing reagent (RNAlater), so the kit have to be able to deal with that? If not, we need suggestions on alternatvie storing of biopsies before extraction - as transfering them directly onto ice is not possible.
Hi Marianne,
I would check out IsoHelix Xtreme DNA isolation kits. The buffer used in the extraction keeps samples shelf stable for a while and might be a good alternative to RNAlater.
https://isohelix.com/products/dna-buccal-swab-isolation/xtreme-dna-isolation-kit/
For DNA isolation, RNAlater is already a better option (than other buffers) if it is available with you. RNAlater would not cause any negative effect on extraction/quality/quantity of DNA. However, if you think it can, just rince the tissue samples with ethanol (but I don't think it would be necessary).
For kits, I would suggest you to go through some of your reference articles and look for the kits they have used. Because more or less, all kits works same/similar in the DNA extraction itself. However, here the main concern would be the removal of eukaryotic DNA. Or if you want to do a microbes specific amplicon sequencing, I don't think it would be a big problem. But since the biopsy will have a huge amount of the eukaryotic DNA, the DNA concentration (microbial) possibly would not be enough for the PCR1. I would suggest you have to test your protocol of DNA isolation and PCR on test samples before you actually go for real samples. You might need to customize few steps. Actually, it happens in many of the cases that one method might not work as good as other for different type of samples.
Thank you so much for your answers, Katherine Bockrath and Abhijeet Singh ! :-)
I think there are many commercial available protocols for DNA extraction. The key thing will be the lysis process that would need to be optimized. A combination of chemical and physical steps might be granted for efficient DNA extraction and purification. Good luck.
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