We understand you may have questions regarding our sequencing services; read further to learn more about common questions surrounding microbial sequencing.
What is microbial DNA Sequencing?
Microbial DNA sequencing is the process of naming a species by identifying its genetic makeup down to the nucleotides. By knowing a fungal or bacterial sample's gene sequence, our lab can compare that DNA to a database of over 20,000 validated species called MicroSEQ ID. This allows us to confidently and accurately report the identity of the microbe down to the genus and the species.
How does Microchem Laboratory genetically sequence bacterial or fungal isolates?
Microchem genetically identifies microbes through a series of steps: growing a pure culture of the isolate, lysing and extracting the isolate's DNA, performing a Polymerase Chain Reaction to amplify a specific DNA region, performing a second round of PCR to attach fluorescent nucleotides to each nucleotide base, capillary electrophoresis to sequence the DNA, and identifying the species using MicroSEQ ID by comparing the sequenced DNA to a genetic database of microbes. For a more in-depth explanation of the process, please visit our page The Science Behind Microbial Identification Using Genetic Sequencing.
What are common instances businesses need to genetically sequence microbes?
Companies often contact our lab to identify an isolate when there is contamination within their product or facility. Microbe contaminants can be found during the manufacturing process, product development, or within sterile environments. Identifying the species' DNA helps prevent further unwanted microbial growth. Please visit How Microbial Identification Can Benefit Your Company to read more about the advantages of sequencing with Microchem.
What is MicroSEQ and how does it work?
MicroSEQ ID is the computer database our laboratory uses to comparatively analyze microbial DNA. MicroSEQ ID compares the isolate's DNA to over 20,000 validated species and gives the results as a phylogenetic tree. The phylogenetic tree shows which species the isolate's DNA is closely related to and its relatives. To learn more about phylogeny's role in DNA sequencing, click here.
How does fungal and bacterial sequencing differ?
For fungal isolates, the D2 region of the DNA is amplified, whereas bacterial isolates are derived from the 16S rRNA gene sequence. These regions are highly conserved, meaning it does not have significant evolutionary differences, but still has differing functions among various species. This allows for an easy comparison of microbes because clear speciation is seen when there is a change in sequence.
Why do we use 16S rRNA for sequencing bacterial samples?
Genetically sequencing the 16S rRNA gene is considered the gold standard of microbial identification. As described above, the 16S rRNA gene is highly conserved and present in almost all bacteria. It acts as a fingerprint for bacterium and allows for seamless identification when compared to a library of organisms.
How do sequencing results complement test reports?
Genetic microbial identification complements tests reports by having data-backed validation of species. Our lab has extensive experience in identifying microbes through traditional methods, but DNA sequencing gives an exact species identity. Visit our page on the comparison of Traditional Microbial Identification Methods vs. Genetic Sequencing to learn more.