Description
The composition of fecal samples is largely dependent on diet which includes fibers, undigested particles, bilirubin, complex polysaccharides, and lipids. Those compounds impair the sample homogenization, decrease both the quantity and quality of the extracted DNA. The SPINeasy® DNA/RNA Kit for Feces integrates our proprietary inhibitor removal expertise and our new technology for selective binding of DNA and RNA. This kit is especially suitable when the sample is limited or when both genomic and transcriptomic manipulations are needed. This thereby avoids any potential variation in microbial community composition from processing bias.
The SPINeasy® DNA/RNA Kit for Feces allows simultaneous isolation of pure microbiome genomic DNA and RNA from challenging feces types irrespective of their composition. Isolated samples showed no contaminants and were immediately ready for use in downstream applications, including qPCR and RT-qPCR without the need of further inhibitor removal step.
Performance
When used on various feces samples, the SPINeasy® DNA/RNA Kit for Feces delivered higher yield, optimal 260/280 and 260/230 ratio and similar or better integrity than the competitor Q kit, indicating a better extraction performance across a wide range of sample compositions (Figure 1).
Figure 1. The SPINeasy® DNA/RNA Kit for Feces extracted DNA and RNA from various fecal samples with high yield and purity. Eleven fecal sample types were processed using SPINeasy® DNA/RNA Kit and competitor Q kit. Yield and purity ratios (A260/280 and A260/230) were measured by fluorometric quantification. Each dot on the plot represents a single extraction. Representative virtual gels (obtained using Agilent 4150 TapeStation analyses) were presented together with the DNA integrity (DIN) and RNA integrity (RIN) values. The red rectangle indicated the contamination of RNA samples with some high molecular weight DNA.
Unlike competitor Q kit, no cross-DNA contamination was observed in RNA samples extracted with the SPINeasy® DNA/RNA Kit for Feces. As shown in Agilent virtual gel (Figure 1, red rectangle) or agarose gel profile (Figure 2), some RNA samples extracted with the competitor Q kit contained DNA contamination; this highlights the outstanding performance of the SPINeasy® DNA/RNA Kit for Feces as compared to other similar commercial kit on the market.
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Figure 2. No DNA contamination in the RNA samples extracted with the SPINeasy® DNA/RNA Kit for Feces. RNA samples extracted from horse, duck and pig fecal samples were visualized on agarose gel. Unlike competitor Q, no smeared nucleic acid corresponding to DNA contamination could be observed in RNA samples extracted with MP Biomedicals’ kit. |