Please share your recommendations for isolating microbial gDNA from faeces and saliva. I'm initially working with cattle but will also be working with clinical samples. I'm having difficulty with matrix inhibitory affects. Downstream application isothermal and PCR amplification. Furthermore, I'm experiencing inhibition from transport media - suitable compatible options for cat 3 organisms? Many thanks in advance
Hi, there are many commercial silica based column kits dedicated to DNA extraction from stool such as QIAamp DNA Stool Mini Kit or NucleoSpin DNA Stool. In homemade methods, β-Mercaptoethanol is added most of the time in extraction buffers and is a strong reducing agent to clean inhibitors. The attached paper compares published methods on DNA extraction from bovine faeces.
For saliva samples, you can use normal columns (QIAGEN, M-NAGEL) as we do for swabs without any problem.
please what of the microbial DNA extraction kit from culture.
Dear Abubakar Abdullahi Lema ,
If you have a pure culture, just dilute some colonies in water (NaCl 9‰) and heat at 95°C for 5 minutes. That all!
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