Hello,
I am starting with the study of vaginal microbiome. We have collected vaginal swabs, centrifugated them (6000 rpm, 10 min) and collected the pellets for the analysis.
I was thinking about using PureLink Microbiome DNA Purification kit from invitrogen. Do you have any experience with this kind of sample and with this kit? Any suggestions?
I had also in mind the QIAamp DNA Microbiome kit from QIAGEN, but the cost is more than double. Any other kit do you recommend me to use?
Thank you very much,
Andrea
Even using a plant DNA extraction kit will give you good yield, as well as maximal removal of inhibitors. Plant kits are good at lysing even the microbes with hardiest cell walls.
best wishes
Try this one.
http://www.tiangen.com/en/?productShow/t1/4/id/8.html
This is a daughter company from Qiagen, but prices are much lower.
Dear Andrea,
if you want to work in the field of microbiome than I would suggest you either use a commercial extraction kit or a nucleic acid extraction machine because you need DNA of good quantity and purity. And you need very good standardization. The costs of a commercial extraction kit are neclectable compared to the costs of a microbiome PCR analysis kit.
Best wishes
Dirk
Thank you very much for your answers. I will finally try with this kit from invitrogen:
https://www.thermofisher.com/es/es/home/life-science/dna-rna-purification-analysis/dna-extraction/microbiome-dna-extraction.html
The thing is that I am not sure about including a previous incubation with lisozyme (most studies include it) or not. They only recommend it for stool samples so I think I will try without it.
Thank you again for your advice.
Best,
Andrea
PureLink microbiome kit works well. or for higher throughput there are MagMax kits with protocols for vaginal swabs
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