Hi,
As the mucin can be the PCR inhibitor, I want to remove (hydrolyze) the mucin from the sputum sample.
I tried to find the product "Mucinase", but nothing came out.
I want to know what product is used to remove (hydrolyze) the mucin.
Thanks.
Hi Jang,
We use Sputolysin® dithiothreitol (DTT) Reagent, CAS 3483-12-3. You can buy it on Sigma-Aldrich ref 560000.
María Luisa Caballero Thank you for your kind reply. Then, how much DTT do I need to add to the sample? My sample volume is about 30ml.
I use a solution 0.1% daily prepared, and mix it 50% v/v with the sample.
María Luisa Caballero Thanks! After mixing the sample with 0.1% DTT solution, I plan to incubate in RT with mild shaking for 30-60min. Lastly, I will extract the DNA from the sample by using DNA extraction kit (QIAgen DNeasy Blood&Tissue kit) and perform 16S PCR. Does it make sense?
Yes, it seems correct to me, but the incubation with the DTT solution, I do it with not gentle shaking, better a little stronger shaking to help disintegration.
María Luisa Caballero Can I ask more questions about mucin? I have some problems with it.....
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