Microbial ecology study – real-time polymerase chain reaction (PCR)
The StepOne™ Real-Time PCR System is a 48-well Real-Time PCR instrument with simple and flexible instrument set-up and usage. It is designed to deliver precise, quantitative Real-Time PCR results for a variety of genomic research applications.
In our previous studies, the StepOne™ Real-Time PCR System was mainly used to quantify the bacteria involved in nitrogen cycle such as ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB) and denitrifiers by amplifying their amoA, NSR and nirK genes respectively. DNA from the soil/media sample was extracted by using a SoilMaster® DNA Extraction Kit (EPICENTRE). The extracted DNA was finally resuspended in 300 uL TE Buffer. Real-time PCR quantification was performed to amplify the target genes, which are in direct proportion of the number of bacteria.
DNA Gel electrophoresis is a method to sort molecules based on their size and charge. It is performed after amplification of DNA via PCR and further used as a preparative technique prior to use of cloning and qPCR. Shorter molecules move faster than longer ones within the agarose gel. DNA can be visualized with ethidium bromide (EB) under ultraviolet light.
Ecotoxicity testing – Algal toxin
Cyanobacteria and the hepatotoxic microcystins were present in 15 central Florida, USA, stormwater ponds where all had detectable levels of microcystin with concentrations ranging from 0.04 to 1.6 μg/L. Cyanobacteria may proliferate in stormwater ponds; cyanotoxins produced by these organisms represent potential public health concerns. Microcystins and Cylindrospermopsin have been found in fresh water throughout the world. Acute poisoning of humans and animals constitutes the most obvious problem from toxic cyanobacterial blooms, and in several cases has lead to death. These toxins mediate their toxicity by inhibiting liver function. The WHO has proposed a provisional upper limit for microcystin-LR of 1.0 ppb (ug/L) in drinking water and a tolerable daily intake (TDI) of cylindrospermopsin as 0.02 g/Kg body weight/day.
Algal toxin was quantitatively analysed using commercially available ELISA kits (Microcystin-ADDA and Cylindrospermopsin kits, Abraxis, LLC, Warminster, PA). The detection limit for each ELISA is 0.1 μg/L (MCLR) and 0.04 μg/L (CYL).
Lake, streams, and wetland restoration Best Management Practices (BMP)
The excess ions such as calcium, iron, sulfate, etc. into the lake have the potential to impact the biological and chemical environment of the lake water. Our study is to explore the interactions between the water column, pore water and the bottom sediment, which compounds the biogeochemical cycle due to the dynamics of phosphorus concentrations in terms of adsorption, desorption, and diffusion processes. This research presented a systematic assessment based on explored the sensitivity with respect to the chemical equilibrium between lake water and lake sediment.
A glove box is a sealed container that allows one to manipulate items where a very high purity inert atmosphere is desired. It is also possible to use a glovebox for manipulation of objects in a vacuum chamber. In our previous study, the glove box was used to provide an anaerobic condition for the measurement of Oxido-reduction Potential (ORP) by filling the glove box with nitrogen gas.