The geomicrobiology lab is primarily used to analyze samples gathered from various deep-sea locales. We carry out extraction and amplification of DNA, identification and cultivation of microorganisms and quantification and characterization of diverse microbial groups in geological and biological samples.
Extraction of genetic material
Extracting genetic material is crucial for addressing research questions involving geomicrobiology. We rely on extracted DNA to quantify and characterize the life forms present in samples originating from a wide range of terrestrial and marine environments. To isolate DNA, we use thoroughly tested extraction kits and protocols customized to each task. Extraction is routinely followed by quantification and visualization of DNA for quality control before further analyses.
Genetic material isolated from environmental samples is often limited in quantity and can be poor in quality. In order to replicate and produce more DNA for planned analyses, the geomicrobiology lab is equipped with Applied Biosystems Venti thermal cyclers, which allow us to carry out Polymerase chain reactions (PCR). This cornerstone molecular biology method allows the rapid production of specific stretches of DNA, specified by short “starter” DNA fragments called primers. This production of DNA is driven by a microbial enzyme DNA Polymerase which in higher amounts, allowing the use of the DNA in downstream analyses such as sequencing, cloning or community analysis of the original sample.
A more specialized instrument is also in place for the real-time monitoring of PCR amplification with the use of fluorescent dyes. The Applied Biosystems StepOnePlus quantitative PCR (qPCR) machine allows for the quantitative assessment of microbiological samples based on the different primers used for amplification.
Cultivation of microorganisms is a classical microbiological approach. In order to better understand deep-sea microorganisms and microbial communities, being able to isolate pure cultures to describe their features and observe their growth can be very useful. To allow for this, the geomicrobiology lab hosts equipment required in the cultivation of diverse microorganisms. Laminar flow benches and fume hoods allow for clean preparation of required growth media, and we have several static and shaking incubators from Binder, Termaks and Gerhardt that facilitate the incubation of chosen microorganisms. We also have a Whitley A35 anaerobic workbench for working with organisms that do not tolerate the presence of oxygen.
Fluorescence in situ Hybridization
The geomicrobiology lab is equipped with a Zeiss Fluorescence microscope. In addition to its high-quality image capturing capabilities, this microscope can also be used for the fluorescence in situ hybridization (FISH) method. With FISH, we can use fluorescent gene-tags to visually image and identify microorganisms with specific genetic properties and to discriminate between different groups of microorganisms in our samples.
We have recently added a Seal Analytical QuAAtro39 AutoAnalyzer system to our laboratory. The system is mainly used to carry out nutrient analyses of porewater samples taken from various environments, informing researchers about the concentrations of a range of important molecules such as NO3–, NO2–, PO4, and NH4+.