All Sylphium sample sets include an Internal Positive Control (IPC) within the preservation solution, located in the small syringe of the sampling set (SYL009). The IPC serves as an efficiency check for the DNA isolation procedure, ensures the quality and purity of the isolated eDNA, and helps prevent false negative results. This internal control consists of a small piece of synthetic xenobiotic DNA. The selected positive control DNA sequence is not found in the aquatic environment and does not interfere with the detection of target organisms. Additionally, the IPC Quantification Kit can be ordered separately or together with the eDNA Isolation Kit. The IPC is detected through the FAM channel.
Kit properties:
The IPC qPCR quantification kit has:
- High resistance to inhibiting factors. Environmental DNA (eDNA) extracts contain often multiple qPCR inhibiting factors. Normal qPCR master mixes are sensitive to these substances.
- Strong fluorescence signal with low background noise. Isolated environmental samples contain residues of naturally occurring auto fluoresce substances that will interfere with the measurements. Environmental samples needs a strong fluorescence signal from the assay in order not to experience background interference.
- Highest possible sensitivity (1 DNA copy per reaction). Environmental water samples may contain very low amounts of target DNA.
- 100% specificity. Isolated DNA from environmental samples contains billions of DNA fragments from bacteria, protozoa, plants, animals, etc. Not only animals from the same order (fish, amphibians, reptiles, mammals, etc.) must be taken in account during primer/probe design, but all known DNA sequences must be checked for nonspecific binding of the primers and probe.
The full method validation report can be found below under “Relevant documents”
The kit has been developed and optimized for use on eDNA isolates purified using the Sylphium Molecular Ecology eDNA Isolation Kit (#SYL002), although other eDNA isolation methods/kits can be used as well. If you need more information on using the obtained isolates from other methods/kits to get reliable results, send an e-mail to info@sylphium.com.
Kit contents:
- Positive control
- Primer/probe mix (10x) for detection of IPC (FAM dye)
- eDNA qPCR master mix:
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- eTaq DNA polymerase
- eTaq qPCR mix
- PCR water
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The kit contains all materials for 200 reactions of 25µl.
Relevant Documents:
- Manual and technical specifications
- Instructions manual (Universal protocol of eDNA qPCR master mix)
- Method validation report
- Lot validation report:
