The genomic DNA from the four microalgal cultures was isolated using hexadecyl-trimethyl-ammonium bromide (CTAB) method with minor modifications [54 ]. The quantity and quality of the DNA were determined using a Nano drop Fluorometer at OD260/280nm for DNA. The 18S rDNA gene of each isolate was amplified using the universal primers; forward primer CV1 (5’-TACCTGGTTGATCCTGCCAGTAG–3′) and reverse primer CV2 (5′- CCAATCCCTAGT CGGCATCGT–3′). For PCR amplification, a reaction mixture (20 μL), containing 50 ng DNA template (2 μL), 1X Taq buffer (2 μL), 0.2 mM of each dNTP mixture (2 μL), 1 μM of forward primer (0.5 μL), 1 μM of reverse primer (0.5 μL), 1.5 mM MgCl2 (1 μL) and 2 Units of Taq DNA polymerase (0.5 μL) (Bangalore Genei, India), and distilled water (11.50 μL), was prepared. PCR amplification was carried out in a Thermocycler (Quanta Bio, USA) with the initial denaturation temperature of 95 °C for 5 min followed by 35 cycles at 95 °C for 30 s, annealing at 45 °C for 30 s, extension at 72 °C for 2 min with a final extension at 72 °C for 10 min. Sequence analysis was then performed on PCR products using BigDye® Terminator kit on ABI 3730XL automatic DNA sequencer (Applied Biosystems). Bioedit v.7.2.0 was used to assemble the 18S rDNA gene sequences. A phylogenetic tree was constructed using the Neighbor-Joining (NJ) method [55 (link)] as implemented in the program MEGA v.6.0 with bootstrapping at 1000 replicates. Microalgae included in the phylogenetic tree were chosen from the NCBI BLAST search.
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Living Beings
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Eukaryote
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Microalgae
Microalgae
Microalgae are microscopic, photosynthetic organisms that thrive in aquatic environments.
These diverse, unicellular species range from green algae to diatoms and cyanobacteria.
Microalgae have garnered significant interest due to their potential applications in biofuels, nutraceuticals, cosmetics, and environmental remediation.
Their rapid growth, ability to fixate carbon dioxide, and production of valuable metabolites make them a promising subject of research and commercialization.
Microalgae's versatility and adaptability offer exciting opportunities for innovatiors to harness their untapped potential and drive breakthroughs in sustainable technology.
Uncover the power of microalgae with PubCompare.ai - your AI-driven platform for effortless protocol discovery and optimization.
These diverse, unicellular species range from green algae to diatoms and cyanobacteria.
Microalgae have garnered significant interest due to their potential applications in biofuels, nutraceuticals, cosmetics, and environmental remediation.
Their rapid growth, ability to fixate carbon dioxide, and production of valuable metabolites make them a promising subject of research and commercialization.
Microalgae's versatility and adaptability offer exciting opportunities for innovatiors to harness their untapped potential and drive breakthroughs in sustainable technology.
Uncover the power of microalgae with PubCompare.ai - your AI-driven platform for effortless protocol discovery and optimization.
Most cited protocols related to «Microalgae»
Buffers
Cetrimonium Bromide
DNA, Ribosomal
Genes
Genome
Magnesium Chloride
Microalgae
Oligonucleotide Primers
Sequence Analysis
Taq Polymerase
Three batches of moribund Crassostrea gigas oysters were collected on the field along the French coasts during abnormal mortality outbreaks in 2008 (Table 1 ). The first batch was composed of oysters collected at Gouville (Normandy - Cotentin), the second of oysters collected in the estuary of the Etel river (Brittany - Morbihan), and the last one corresponded to oysters collected on the site of Aulne located in the Bay of Brest (Brittany - Finistère). These animals were checked for OsHV-1 detection and served for the preparation of tissue homogenates. A control tissue homogenate was prepared from the batch of healthy appearing oysters used for experimental assays and showing no mortality in 2008 (see below; Table 1 ).
Experimental infection trials were performed on healthy appearing C. gigas spat (one year old) purchased in November 2008 from a shellfish farm located on the French Mediterranean coast. No mortality event has been reported at this shellfish farm location during 2008. Oyster spat sized around 40 mm in length, with a mean weight of 5 grams. Oysters were placed in the Ifremer's facilities (Laboratoire de Génétique et Pathologie, La Tremblade, France) in a single tank of 200 L containing filtered (1 μm) seawater and slowly acclimated to 22°C increasing the temperature of 1°C per day. During this period, oysters were fed daily by addition of 2 liters of microalgae Skeletonema costatum (1.5 103 cells/mL). Oysters did not present any mortality or other symptom of disease at this time. At the end of the acclimatization period and just before the beginning of the experiment, a set of 20 individuals was assessed by real time quantitative PCR in order to evaluate the initial OsHV-1 DNA detection.
No ethical approval has been requested for the present study because experimental research has been conducted on Pacific oysters (invertebrates). Oysters don't possess a central nervous system.
Experimental infection trials were performed on healthy appearing C. gigas spat (one year old) purchased in November 2008 from a shellfish farm located on the French Mediterranean coast. No mortality event has been reported at this shellfish farm location during 2008. Oyster spat sized around 40 mm in length, with a mean weight of 5 grams. Oysters were placed in the Ifremer's facilities (Laboratoire de Génétique et Pathologie, La Tremblade, France) in a single tank of 200 L containing filtered (1 μm) seawater and slowly acclimated to 22°C increasing the temperature of 1°C per day. During this period, oysters were fed daily by addition of 2 liters of microalgae Skeletonema costatum (1.5 103 cells/mL). Oysters did not present any mortality or other symptom of disease at this time. At the end of the acclimatization period and just before the beginning of the experiment, a set of 20 individuals was assessed by real time quantitative PCR in order to evaluate the initial OsHV-1 DNA detection.
No ethical approval has been requested for the present study because experimental research has been conducted on Pacific oysters (invertebrates). Oysters don't possess a central nervous system.
Acclimatization
Animals
Biological Assay
Cells
Central Nervous System
Crassostrea gigas
Disease Outbreaks
Estuaries
Infection
Invertebrates
Microalgae
Oysters
Real-Time Polymerase Chain Reaction
Rivers
Shellfish
Tissues
Three bacterial cultures with a broad range of genomic G+C content, for which finished genomes are available in public databases, were employed in the optimization of WGA-X: Prochlorococcus marinus, Eschericia coli, and Meiothermus ruber (Supplementary Table 1 ). Cultures of two marine unicellular algae, Thalassiosira pseudonana and Ostreococcus lucimarinus (Supplementary Table 1 ) were also used in the subsequent benchmarking of optimized WGA-X and its comparison to MDA.
For marine prokaryote and extracellular particle analyses, the Gulf of Maine surface water was collected from 1 m depth in Boothbay Harbor, Maine (43°50′39.87″ N, 69°38′27.49″ W) on 15 June, 2011. One ml aliquots were amended with 5% glycerol and 1× TE buffer (all final concentrations), and stored at −80 °C until further analysis. The marine microalgae sample was collected from the same location on 16 September, 2009 and cryopreserved with 6% glycine betaine and 1× TE buffer (all final concentrations) at −80 °C. The soil sample was collected from 0–10 cm depth in a residential garden in Nobleboro, Maine (44°5′48.10″ N, 69°29′10.56″ W) on 5 May, 2015. Approximately 5 g of the soil sample were mixed with 30 ml sterile-filtered PBS, vortexed for 30 s at maximum speed, and centrifuged for 30 s at 2000 rpm (800×g). The obtained supernatant was used for cell sorting within 30 min, and processed as described above.
For marine prokaryote and extracellular particle analyses, the Gulf of Maine surface water was collected from 1 m depth in Boothbay Harbor, Maine (43°50′39.87″ N, 69°38′27.49″ W) on 15 June, 2011. One ml aliquots were amended with 5% glycerol and 1× TE buffer (all final concentrations), and stored at −80 °C until further analysis. The marine microalgae sample was collected from the same location on 16 September, 2009 and cryopreserved with 6% glycine betaine and 1× TE buffer (all final concentrations) at −80 °C. The soil sample was collected from 0–10 cm depth in a residential garden in Nobleboro, Maine (44°5′48.10″ N, 69°29′10.56″ W) on 5 May, 2015. Approximately 5 g of the soil sample were mixed with 30 ml sterile-filtered PBS, vortexed for 30 s at maximum speed, and centrifuged for 30 s at 2000 rpm (800×g). The obtained supernatant was used for cell sorting within 30 min, and processed as described above.
Betaine
Buffers
Genome
Genome, Bacterial
Glycerin
Marines
Meiothermus ruber
Microalgae
Prochlorococcus marinus
Prokaryotic Cells
Strains
For a proper measurement of the antioxidative potential of the tested microalgae using the DPPH radical, the following procedure was successfully executed. For choosing the suitable wavelength, wavelength scans from 440 to 750 nm were done for both measuring and reference cuvette. The scans were run against pure methanol. Measurements in the measuring cuvette were performed 30 min after addition of DPPH in order to give enough time for the reaction of the cellular antioxidants with DPPH. During this 30 min, reference and measuring cuvette were kept in the dark at ambient temperature. Additionally, the reference cuvette was measured before and after the reaction in the measuring cuvette had taken place for verification that no changes of the absorptive properties of the microalgae extract had occurred in the absence of DPPH.
After wavelength scans were done (3 min) 0.3 mL of the α-tocopherol solution was added to both measuring and reference cuvette in order to achieve full reduction of the DPPH radicals. Then, the wavelength scans were repeated. This served to select an adequate wavelength which is influenced only by the DPPH radical (here 550 nm).
The absorbance decrease was then measured by using measuring and reference cuvette prepared and treated in the same way described above. But here the absorbance of the measuring cuvette was measured at 550 nm against the reference cuvette for elimination of the absorptive properties of the microalgae extract.
All measurements were performed in triplicate. Here, averages and standard deviations are presented.
After wavelength scans were done (3 min) 0.3 mL of the α-tocopherol solution was added to both measuring and reference cuvette in order to achieve full reduction of the DPPH radicals. Then, the wavelength scans were repeated. This served to select an adequate wavelength which is influenced only by the DPPH radical (here 550 nm).
The absorbance decrease was then measured by using measuring and reference cuvette prepared and treated in the same way described above. But here the absorbance of the measuring cuvette was measured at 550 nm against the reference cuvette for elimination of the absorptive properties of the microalgae extract.
All measurements were performed in triplicate. Here, averages and standard deviations are presented.
alpha-Tocopherol
Antioxidants
Cells
Methanol
Microalgae
Radionuclide Imaging
Strains of the green microalgae B. braunii SAG 30.81, S. obliquus SAG 276-3a, N. conjuncta SAG 78.80, N. terrestris UTEX B. 947, and N. texensis SAG 99.80 originating from the SAG Culture Collection of Algae and the UTEX Culture Collection of Algae were inoculated from solid into sterile liquid Kessler’s medium to obtain a sufficiently large quantity of algal biomass required for the experiments. Preliminary semi-continuous cultures were run under light (Osram L58W/765 cool daylight) and temperature 24 ± 1 °C for 60 days until biomass suitable for the experiments was obtained. The intensity of photosynthetically active light (PPFD) was 60 μmol m−2 s−1.
The biomass obtained was used (1) for determination of the relationship curves between the optical density of algal culture measured with the spectrophotometric method (Unicam Helios, UK) at the 650-nm wavelength and the dry weight (determined with the weighing method) of algae growing under the conditions specified above, and (2) as an inoculum for the growth experiments.
The biomass obtained was used (1) for determination of the relationship curves between the optical density of algal culture measured with the spectrophotometric method (Unicam Helios, UK) at the 650-nm wavelength and the dry weight (determined with the weighing method) of algae growing under the conditions specified above, and (2) as an inoculum for the growth experiments.
Light
Microalgae
Spectrophotometry
Sterility, Reproductive
Strains
Vision
Most recents protocols related to «Microalgae»
Example 6
Strain 5 was subjected to another round of mutagenesis with increasing concentrations and exposure time to 4-NQO (37 μM for 30 minutes at 28° C.). This population of cells was subsequently subdivided and grown in standard lipid production medium supplemented with a range of cerulenin concentrations (7-50 μM). Cells from all concentrations were pooled and fractionated over a 60% Percoll/0.15 M NaCl density gradient. Oil laden cells recovered from a density zone of 1.02 g/mL were plated and assessed for glucose consumption and fatty acid profile. One of these clones was subsequently stabilized and given the strain designation “Strain 6”.
Cells
Cerulenin
Clone Cells
Fatty Acids
Glucose
Lipids
Microalgae
Mutagenesis
Oleic Acid
Percoll
Sodium Chloride
Strains
Triglycerides
Example 5
A subpopulation of the Strain 4 lineage was concurrently subjected to an enrichment strategy employing one round of enrichment in an inhibitor cocktail (Inhibitor Cocktail 2 in
Cardiac Arrest
Cells
Clomiphene Citrate
Clone Cells
Fatty Acids
Fluphenazine
Glucose
Microalgae
Oleic Acid
Population Group
Strains
Terfenadine
Triglycerides
Triparanol
Example 4
As an alternative to stabilizing Strain 3, a new round of mutagenesis was pursued for Strain 2 utilizing treatment of cells with the mutagen 4-nitroquinoline-1-oxide (4-NQO) for 5 minutes at 28° C. Mutagenized cells were enriched by growing under conditions of limited glucose (14 g/L) for three days, then the cells were subjected to fraction over a 60% Percoll/0.15 M NaCl density gradient. Cells recovered from a density zone of 1.06 g/mL were plated and assessed for glucose consumption and fatty acid profile. One of these clones was subsequently stabilized and given the strain designation “Strain 4”.
Cells
Clone Cells
Cultured Cells
Fatty Acids
Glucose
Microalgae
Mutagenesis
Mutagens
Oleic Acid
Oxides
Percoll
Sodium Chloride
Strains
Triglycerides
Pyrolysis
experiments of the microalgal samples before and after the addition
of the catalyst were performed via thermogravimetric analysis (TGA)
in nitrogen and air using the Setaram TAG 16 Simultaneous Symmetrical
Thermo Analyzer (Setaram, France). First, 10–20 mg of sample
in an alumina cup was exposed to nitrogen (140 mL/min) for 10 min
before being heated from room temperature to 850 °C at a 10 °C/min
heating rate. This temperature was maintained in the nitrogen gas
stream for 20 min before switching to air at a flow rate of 70 mL/min
for another 20 min at the same temperature. The temperature was then
reduced to 20 °C at a rate of 20 °C/min in an air stream
(70 mL/min). This temperature was held constant for 30 min.
The TG and dTG curves of the pyrolysis of Botryococcus braunii with and without the catalyst were analyzed based on the method
described in our previous study.39 (link) The
curves were divided into three temperature ranges in a manner similar
to that reported by Chen et al.53 (link) The temperature
at which the first peak of the dTG curve slopes is the maximum temperature
in the first stage (lowest dTG value). The first stage of the dTG
curve has only one peak, while the second stage has several peaks.
The temperature range in the second stage is the temperature at which
the peaks on the dTG curve have flattened after the first stage is
completed. The temperature after the end of the second stage is the
initial temperature in the third stage.
experiments of the microalgal samples before and after the addition
of the catalyst were performed via thermogravimetric analysis (TGA)
in nitrogen and air using the Setaram TAG 16 Simultaneous Symmetrical
Thermo Analyzer (Setaram, France). First, 10–20 mg of sample
in an alumina cup was exposed to nitrogen (140 mL/min) for 10 min
before being heated from room temperature to 850 °C at a 10 °C/min
heating rate. This temperature was maintained in the nitrogen gas
stream for 20 min before switching to air at a flow rate of 70 mL/min
for another 20 min at the same temperature. The temperature was then
reduced to 20 °C at a rate of 20 °C/min in an air stream
(70 mL/min). This temperature was held constant for 30 min.
The TG and dTG curves of the pyrolysis of Botryococcus braunii with and without the catalyst were analyzed based on the method
described in our previous study.39 (link) The
curves were divided into three temperature ranges in a manner similar
to that reported by Chen et al.53 (link) The temperature
at which the first peak of the dTG curve slopes is the maximum temperature
in the first stage (lowest dTG value). The first stage of the dTG
curve has only one peak, while the second stage has several peaks.
The temperature range in the second stage is the temperature at which
the peaks on the dTG curve have flattened after the first stage is
completed. The temperature after the end of the second stage is the
initial temperature in the third stage.
ARID1A protein, human
Microalgae
Nitrogen
Oxide, Aluminum
Pyrolysis
Botryococcus braunii samples were isolated
from the fresh waters of Tenggarong, Kutai Kartanegara Regency, East
Kalimantan, Indonesia. The green microalgae were then cultivated at
the Laboratory of Animal Physiology, Developmental and Molecular Animals,
Faculty of Mathematics and Natural Sciences, Mulawarman University,
Indonesia, using the method described in our previous paper.39 (link) After harvesting, the algal paste was centrifuged
for 30 s at a speed of 5000 rpm. Following that, the algal samples
were homogenized for 30 s at a speed of 300 rpm and dried for 24 h
in a freeze-dryer. The freeze-dried samples were kept at 20 °C.
The water content, ash content, protein content, total chlorophyll,
and fatty acid composition of the freeze-dried samples were analyzed
by the methods described in our previous study.39 (link) The physical and chemical properties of Botryococcus
braunii are presented inTable 1 .
from the fresh waters of Tenggarong, Kutai Kartanegara Regency, East
Kalimantan, Indonesia. The green microalgae were then cultivated at
the Laboratory of Animal Physiology, Developmental and Molecular Animals,
Faculty of Mathematics and Natural Sciences, Mulawarman University,
Indonesia, using the method described in our previous paper.39 (link) After harvesting, the algal paste was centrifuged
for 30 s at a speed of 5000 rpm. Following that, the algal samples
were homogenized for 30 s at a speed of 300 rpm and dried for 24 h
in a freeze-dryer. The freeze-dried samples were kept at 20 °C.
The water content, ash content, protein content, total chlorophyll,
and fatty acid composition of the freeze-dried samples were analyzed
by the methods described in our previous study.39 (link) The physical and chemical properties of Botryococcus
braunii are presented in
Animals
Animals, Laboratory
chemical properties
Chlorophyll
Faculty
Fatty Acids
Freezing
Microalgae
Paste
Physical Examination
physiology
Proteins
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DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.
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Whatman No. 1 filter paper is a general-purpose cellulose-based filter paper used for a variety of laboratory filtration applications. It is designed to provide reliable and consistent filtration performance.
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Methanol is a clear, colorless, and flammable liquid that is widely used in various industrial and laboratory applications. It serves as a solvent, fuel, and chemical intermediate. Methanol has a simple chemical formula of CH3OH and a boiling point of 64.7°C. It is a versatile compound that is widely used in the production of other chemicals, as well as in the fuel industry.
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Sea salts are a type of natural salt derived from the evaporation of seawater. They are used in various laboratory applications as a source of minerals and trace elements.
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GF/C filters are laboratory filter papers designed for general filtration purposes. They are made of glass microfibers and have a nominal pore size of 1.2 microns. GF/C filters are suitable for a variety of filtration applications, including sample preparation, particle retention, and liquid-solid separation.
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The UV-1800 is a UV-Visible spectrophotometer manufactured by Shimadzu. It is designed to measure the absorbance or transmittance of light in the ultraviolet and visible wavelength regions. The UV-1800 can be used to analyze the concentration and purity of various samples, such as organic compounds, proteins, and DNA.
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The Axio Observer A1 is a compact and versatile inverted microscope designed for routine observation and imaging. It features a stable frame, adjustable viewing angle, and a range of objectives to accommodate various sample types. The Axio Observer A1 provides a straightforward and efficient solution for basic microscopy applications.
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Hydrochloric acid is a commonly used laboratory reagent. It is a clear, colorless, and highly corrosive liquid with a pungent odor. Hydrochloric acid is an aqueous solution of hydrogen chloride gas.
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The CytoFLEX flow cytometer is a versatile instrument designed for high-performance cell analysis. It utilizes advanced optical and fluidic technologies to enable accurate and reliable detection and enumeration of a wide range of cell populations. The CytoFLEX is capable of simultaneous multi-parameter analysis, providing researchers with comprehensive data on cellular characteristics and populations.
More about "Microalgae"
Microalgae, also known as phytoplankton or microphytes, are tiny, photosynthetic organisms that thrive in aqueous environments.
These diverse, single-celled species range from green algae to diatoms and cyanobacteria (blue-green algae).
Microalgae have garnered significant interest due to their potential applications in biofuels, nutraceuticals, cosmetics, and environmental remediation.
Their rapid growth, ability to fix carbon dioxide, and production of valuable metabolites make them a promising subject of research and commercialization.
Microalgae's versatility and adaptability offer exciting opportunities for innovators to harness their untapped potential and drive breakthroughs in sustainable technology.
Researchers often utilize various tools and techniques to cultivate and analyze microalgae, such as DMSO (dimethyl sulfoxide) as a solvent, Whatman No. 1 filter paper for filtration, and methanol for extraction.
Sea salts provide the necessary minerals for microalgae growth, while GF/C filters are used to collect biomass.
Spectrophotometers like the UV-1800 and microscopes like the Axio Observer A1 are employed to measure and visualize microalgal cells.
Acidic compounds, such as hydrochloric acid, may be used for pH adjustment, and plate readers like the Synergy H1 can analyze microalgal metabolism and growth.
Flow cytometry, using instruments like the CytoFLEX, enables the rapid quantification and sorting of individual microalgal cells, providing insights into their physiological state and population dynamics.
Discover the power of PubCompare.ai, your AI-driven platform for effortless protocol discovery and optimization, to accelerate your microalgae research and unlock the full potential of these remarkable organisms.
These diverse, single-celled species range from green algae to diatoms and cyanobacteria (blue-green algae).
Microalgae have garnered significant interest due to their potential applications in biofuels, nutraceuticals, cosmetics, and environmental remediation.
Their rapid growth, ability to fix carbon dioxide, and production of valuable metabolites make them a promising subject of research and commercialization.
Microalgae's versatility and adaptability offer exciting opportunities for innovators to harness their untapped potential and drive breakthroughs in sustainable technology.
Researchers often utilize various tools and techniques to cultivate and analyze microalgae, such as DMSO (dimethyl sulfoxide) as a solvent, Whatman No. 1 filter paper for filtration, and methanol for extraction.
Sea salts provide the necessary minerals for microalgae growth, while GF/C filters are used to collect biomass.
Spectrophotometers like the UV-1800 and microscopes like the Axio Observer A1 are employed to measure and visualize microalgal cells.
Acidic compounds, such as hydrochloric acid, may be used for pH adjustment, and plate readers like the Synergy H1 can analyze microalgal metabolism and growth.
Flow cytometry, using instruments like the CytoFLEX, enables the rapid quantification and sorting of individual microalgal cells, providing insights into their physiological state and population dynamics.
Discover the power of PubCompare.ai, your AI-driven platform for effortless protocol discovery and optimization, to accelerate your microalgae research and unlock the full potential of these remarkable organisms.