Hello everyone,
I would like to ask you some questions about extracting human RNA from stool samples and performing qRT-PCR.
1. When extracting RNA, how to collect as much human RNA as possible? Since feces contain a lot of bacteria, even if we try to filter, centrifuge, etc., we often get bacterial RNA. We also tried immunomagnetic beads to capture intestinal epithelial cells in feces, but the recovery rate was still unsatisfactory.
2. How to remove inhibitors in qRT-PCR reaction when extracting RNA? Although some fecal total RNA extraction kits have stated that they can effectively remove inhibitors in qPCR reactions, since our goal is a small amount of human RNA in total RNA, we hope to increase the amount of RNA input in the reaction system, but at the same time An inhibitory effect is not expected.
Hope you can give your valuable opinions.
Thanks!
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