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DNA Kit

How to detect DNA from the Exosome Standards?

Vesicles usually contains a very small amount of DNA, the most of it is fragmented and single stranded, and the quantity is not enough for an agarose gel analysis. DNA can be quantify by nanodrop analysis and by qPCR.  

Is there any possibility of genomic DNA or mitochondrial DNA (mtDNA) contamination using EXO-DNA Extraction Kits?

EXO-DNA Extraction Kits allows the isolation of circulating+extracellular vesicle (EV)-associated DNA and in EVs is contained genomic DNA. It is also possible the presence of some mitochondrial DNA in EVs, but the evidences about this issue are not still very clear. If you want to limit the analysis only to the DNA contained in EVs, it is sufficient to make a DNase treatment after the precipitation of vesicles+circulating DNA, before to add the lysis buffer. Rather, vesicles protect the DNA from the DNase treatment, which eliminates only the circulating DNA fraction.

Is there any possibility of ribosomal RNA contamination using ExoDNA™ Extraction Kits?

Yes, there is a possibility of RNA contamination, since the vesicles contain RNA, in particular miRNAs. Percentage of ribosomal RNA is very small in vesicles. Presence of RNA does not cause problems since it is very quickly degraded. It is possible to make an optional treatment with RNase to eliminate the possible contaminations.

Does EXO-DNAExtraction Kits adopt RNase treatment step?

No, the ExoDNA™ Extraction Kits does not adopt any RNase treatment, but it is possible to do the RNase treatment once the DNA is eluted.

Regarding the lyophilized exosome standards, do you also the characterize their DNA/RNA contents too in addition to the particle number and the protein content?

Lyophilized standards are currently used as a positive control in RNA and DNA kit, so it is possible to use them for isolation of nucleic acids, although each lot has characterized for protein content and particle number (NTA). In addition, many customers are using as mutation control in spike-in experiments.