Intended Use
For the enumeration of enterococci in water and other liquids by the membrane filtration technique.
Principle and Interpretation
The growth of the entire accompanying Gram-negative microbial flora is inhibited by sodium azide. Enterococci reduce 2,3,5-Triphenyl tetrazolium chloride (TTC) to give a red formazan inside the bacterial cell, their colonies are thus red. Nitrogen, minerals, and amino acids are provided by the tryptose whilst yeast extract supplies vitamins. Glucose acts as the carbon source, dipotassium phosphate buffers the medium, and agar-agar is the solidifying agent.
Formulation
| Ingredients | /liter |
| Tryptose | 20.0g |
| Yeast extract | 5.0g |
| Glucose | 2.0g |
| Disodium hydrogen phosphate | 4.0g |
| Sodium azide | 0.4g |
| 2,3,5-Triphenyl Tetrazolium chloride | 0.1g |
| Agar | 10.0g |
| pH 7.2±0.1 at 25°C | |
Preparation
Dissolve 41.5 g in 1 L of purified water. Heat in boiling water and agitate frequently until completely dissolved. Sterilize by further heat for 20 minutes in the boiling water bath.
Quality Control
Cultural characteristics observed after incubation at 34-38°C for 40-48hours
| Quality control strains | Growth | Colony color |
| Enterococcus faecalis ATCC29212 | Good growth,PR≥0.5 | deep red coloured colonies |
| Escherichia coli ATCC25922 | Total inhibition | – |
Sorage and Shelf Life
Keep container tightly closed, store in a cool, dry place, away from bright light. Storage period of 3 years.
Precautions
1. When weighing the dehydrated medium, please wear masks to avoid causing respiratory system discomfort
2. Keep container tightly closed after using to prevent clumping.
Waste Disposal
Microbiological contamination was disposed by autoclaving at 121°C for 30 minutes.
Intended Use For the enumeration of enterococci in water and other liquids by the membrane filtration technique. Principle and Interpretation The growth of the entire accompanying Gram-neg……
DBCO-PEG12-Maleimide is a PEG linker containing a DBCO moiety and a terminal primary maleimide group. The maleimide group will react with a thiol group to form a covalent bond, enabling the connection of biomolecule with a thiol. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
DBCO-PEG12-Maleimide is a PEG linker containing a DBCO moiety and a terminal primary maleimide group. The maleimide group will react with a thiol group to form a covalent bond, enabling the connection of biomolecule with a thiol. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Not all cyanobacterial strains produce toxins. However, the toxin-producing strains cannot be distinguished from the nontoxin-producing strains by traditional light microscopy, commonlyused to monitor water bodies. An alternative for the differentiation of potentially toxic strains from nontoxic strains is to use molecular methods to detect the presence of toxin biosynthetic genes. Such methods are already available and could be used for the detection and identification of potential microcystin and nodularin producers present in environmental samples (Attogene catalog number NA2024).
Screening for the toxin itself, can be very costly. In turn, real time PCR for the detection of a gene region responsible for assembling in cyanobacterial strains and environmental samples can be a key indicator for the prescense of cyanobacteria capable of expressing the aetokthonotoxin toxin. Attogen has thus, designed primer pairs and probes targeting a the conserved gene region in order to enable the amplification and detection of several producer genera using real time PCR. Screening for the toxin genes can save significant costs and act as a triage for samples needing to be analyzed for the toxin itself.
Cyanobacterial neurotoxin aetokthonotoxin (AETX), a peculiar pentabrominated biindole alkaloid implicated in fatal Vacuolar Myelinopathy. This neurodegenerative disease was first recorded in 1994 during an outbreak of bald-eagle poisonings at De Gray Lake in Arkansas, USA. AETX was experimentally confirmed to be produced by the true branching heterocytous cyanobacterium Aetokthonos hydrillicola. The production of AETX is dependent on bromide (Br−) availability, and likely linked to its hyper-accumulation by the host plan. Thus regular monitoring of A. hydrillicola (accompanied by assessment of Br− and AETX levels) is highly advisable to predict the possible threat of further VM outbreaks.
The cyanobacterial AetA gene which encodes the unique FAD-dependent halogenase involved in the pathway for AETX synthesis has been adapted to develop a -aetokthonotoxin specific quantitative PCR (qPCR) assay.
Real time qPCR kit for AetA gene
For screening aetokthonotoxin gene cluster
Use in combination with Attogene Algae DNA isolation kit