Our product range includes a wide range of ammonium persulphate hi ar acs, antibiotic assay medium no. 36, antibiotic assay medium no 32, antibiotic assay medium no 36, antibiotic assay medium no 3 assay broth and pseudomonas agar for pyocyanin.
Ammonium persulphate, often labeled "Hi AR" (Hi Media AR or "High AR"), is a white crystalline powder that is highly soluble in water. It's an inorganic compound with the formula (NH4)2S2O8. This reagent is used in various applications like polymer chemistry, as an etchant, and as a cleaning and bleaching agent. Here's a more detailed breakdown:Key Features:
Appearance: White, crystalline powder.
Solubility: Highly soluble in water.
Chemical Formula: (NH4)2S2O8.
Purity: Hi AR typically indicates a high purity standard, often meeting AR (Analytical Reagent) or ACS (American Chemical Society) standards, which means it's suitable for use in analytical chemistry and other applications where high purity is crucial.
Applications:
Polymer Chemistry: Used in the polymerization of various monomers, including aniline, where it acts as an oxidant.
Etching: Used as an etchant in various materials.
Cleaning and Bleaching: Used as a cleaning and bleaching agent in various applications.
Other Uses: It's also used in the preparation of polyacrylamide gels in biochemistry and molecular biology.
Specifications:Ammonium persulphate (Hi AR) typically has specifications that include:
Assay: Minimum 99% by iodometric assay.
Insoluble matter: Maximum 0.005%.
Sulphated ash: Maximum 0.05%.
Acidity (H2SO4): Maximum 0.2%.
Chloride (CI): Maximum 0.0005%.
Iron (Fe): Maximum 0.001%.
Heavy metals (Pb): Maximum 0.001%.
Safety:
Oxidizer: Ammonium persulphate is a strong oxidizing agent.
Irritant: Airborne dust can be irritating to the eyes, nose, throat, lungs, and skin.
Antibiotic Assay Medium No. 36 is a Soya Bean Casein Digest Agar, also known as Tryptone Soya Agar, used primarily for microbiological assays to determine the potency of antibiotics. It serves as a seed agar in antibiotic assays like the agar diffusion assay and is also used for the cultivation of various fastidious and non-fastidious microorganisms, as well as for sterility testing. Key Characteristics:
Composition:Made from Soya Bean Casein Digest, it provides nutrients to support the growth of a broad range of microorganisms, including fastidious ones.
Application in Antibiotic Assays:It's used as a seed agar in the cylinder-plate method to measure the zone of inhibition caused by an antibiotic, and also for turbidimetric assays where growth in liquid culture is measured.
Microbiological Uses:
Cultivation: It supports the growth of a wide variety of microorganisms, including fastidious organisms like Neisseria and Brucella, and non-fastidious ones.
Isolation: Used for isolating microorganisms from various specimens.
Maintenance: Serves as a maintenance medium for stock cultures and specific organisms, such as Mycobacterium smegmatis for bleomycin assays.
Regulatory Compliance:It is recommended by various pharmacopoeias and conforms to United States Pharmacopoeia (USP) standards for use in pharmaceutical procedures.
Inhibitor-Free:Its simple, inhibitor-free composition makes it suitable for detecting antimicrobial agents in products and for other microbial limit tests.
Antibiotic Assay Medium No. 32 is a specialized culture medium used for preparing an inoculum of Bacillus spizizenii to test the potency of dihydrostreptomycin and vancomycin. It is formulated to provide the necessary nutrients and growth factors for the microorganism. This medium is a modified version of Antibiotic Assay Medium No. 1, and is used for plate assays of these antibiotics. Here's a more detailed breakdown:
Purpose:Used to develop an inoculum of Bacillus spizizenii for testing the potency of dihydrostreptomycin and vancomycin.
Key Components:Contains essential nutrients, vitamins, minerals, trace elements, and growth factors supplied by peptone, trytone, yeast extract, and HM peptone B.
Carbon Source:Dextrose is included as a carbon source to stimulate the growth of the test microorganism.
Growth and Sporulation:Manganese sulfate facilitates sporulation and growth of Bacillus subtilis, a common test organism used in these assays.
Formulation:The medium is prepared according to the United States Pharmacopoeia and Food and Drug Administration specifications.
Antibiotic Assay Medium No. 36 is a specialized microbiological medium used for antibiotic assays, particularly for determining the potency of antibiotics. It is a general-purpose medium recommended for cultivating a wide range of microorganisms, including both fastidious and non-fastidious species. This medium is also used for sterility testing in pharmaceutical procedures and is referenced in the US Pharmacopeia. Key Features and Uses:
Antibiotic Assays:It's a standard medium for antibiotic assays, especially for bleomycin, using Mycobacterium smegmatis as the test organism.
Microorganism Cultivation:It supports the growth of various microorganisms, making it suitable for a broad range of applications, including isolation and cultivation.
Sterility Testing:It is used for sterility testing in pharmaceutical settings, ensuring the absence of microbial contamination in products.
Seed Agar:It is often used as a seed agar for agar diffusion assays, a method for determining antibiotic potency.
Pharmaceutical Applications:It is widely used in pharmaceutical manufacturing for quality control and potency determination of antibiotics.
Maintenance of Cultures:It can be used to maintain stock cultures of microorganisms like Pseudomonas aeruginosa for antibiotic testing.
Composition:While the exact formulation can vary slightly between manufacturers, it generally includes a peptone base, dextrose, and other components to support microbial growth. It may also be available in a vegetable-based (Veg) version, replacing animal-derived peptones with vegetable-based hydrolysates. Important Considerations:
Freshly Prepared Plates:For accurate antibiotic assays, freshly prepared plates of Antibiotic Assay Medium No. 36 are recommended.
Sterilization:All materials and media must be properly sterilized before use to prevent contamination.
Storage:Store the dehydrated powder in a dry place, away from direct sunlight, and tightly sealed to prevent moisture absorption.
Antibiotic Assay Medium No. 3, also known as Assay Broth, is a microbiological medium used to determine the potency of antibiotics in pharmaceutical and food products. It's specifically designed for the turbidimetric method, which measures the change or inhibition of growth of a test microorganism in a liquid medium containing varying concentrations of the antibiotic. Here's a more detailed description:Purpose:
Antibiotic Assay:This medium is primarily used for the microbiological assay of antibiotics, a process that determines the strength or effectiveness of an antibiotic.
Turbidimetric Method:The medium is specifically formulated for use in the turbidimetric method, where the growth of a test organism (usually bacteria) is measured by how much light it scatters (turbidity) in the liquid medium.
Quantitative Analysis:By measuring the turbidity, the potency of the antibiotic can be determined by comparing the results to a reference standard.
Composition:
Peptone (Peptic Digest of Animal Tissue): Provides essential nutrients like nitrogen and amino acids for microbial growth.
Beef Extract: Another source of nitrogen and other growth factors.
Yeast Extract: A source of B-complex vitamins, crucial for microbial metabolism.
Dextrose (Glucose): Provides a readily available source of energy for microbial growth.
Sodium Chloride: Maintains osmotic equilibrium and ensures cell viability.
Dipotassium Phosphate and Monopotassium Phosphate: Act as buffering agents to maintain a stable pH.
Function:
1. Nutrient Support:The medium provides the necessary nutrients for the growth of the test microorganism.
2. Growth Enhancement:Components like yeast extract and dextrose are included to promote robust and consistent growth of the test organism.
3. Buffering:The phosphate salts help to maintain a stable pH, which is crucial for the antibiotic's activity and the microorganism's growth.
4. Osmotic Balance:Sodium chloride maintains the appropriate salt concentration, preventing cell damage and ensuring accurate growth measurements.
How it works in a turbidimetric assay:
The antibiotic sample is diluted into different concentrations in the Assay Broth.
The test microorganism is added to each dilution.
The tubes are incubated under controlled conditions.
After incubation, the turbidity (cloudiness) of each tube is measured using a spectrophotometer.
The amount of growth (measured by turbidity) is compared to a standard curve of known antibiotic concentrations to determine the potency of the sample.
Pseudomonas Agar (for Pyocyanin) is a specialized culture medium used to detect and enhance the production of pyocyanin, a blue-green pigment produced by Pseudomonas species, particularly P. aeruginosa. It is a modification of the King's A medium and is used in both clinical and research settings to identify and study these bacteria. Here's a more detailed description:Purpose:
Detection of Pyocyanin:The primary purpose is to facilitate the production and observation of pyocyanin, a pigment that helps in identifying Pseudomonas species, especially P. aeruginosa.
Inhibition of Fluorescein:The medium is formulated to suppress the production of fluorescein (a yellow-green pigment also produced by some Pseudomonas species), which can sometimes mask the blue color of pyocyanin.
Microbial Limit Tests:It's also used in microbial limit tests for pharmaceutical and other biological preparations, as per the U.S. Pharmacopoeia.
Key Components and their roles:
Pancreatic Digest of Gelatin: Provides essential nutrients, amino acids, and trace elements for bacterial growth.
Magnesium Chloride and Potassium Sulfate: These ingredients enhance the production of pyocyanin.
Glycerin: Acts as an energy source, further promoting pyocyanin production.
Agar: Serves as the solidifying agent for the medium.
How it works:
When Pseudomonas species, especially P. aeruginosa, are cultured on Pseudomonas Agar (for Pyocyanin), they are encouraged to produce pyocyanin.
The pigment diffuses into the agar, and the characteristic blue-green color becomes visible, aiding in the identification of the bacteria.
The medium's formulation inhibits the production of fluorescein, preventing it from interfering with pyocyanin detection.
In summary: Pseudomonas Agar (for Pyocyanin) is a valuable tool for microbiology labs, enabling the easy identification of Pseudomonas species through the specific detection of the pyocyanin pigment.
MacConkey Agar Medium 8 is a selective and differential medium used to isolate and differentiate Gram-negative bacteria, particularly enteric pathogens like E. coli and Salmonella. It achieves this by inhibiting Gram-positive bacteria with bile salts and crystal violet, and differentiating lactose-fermenting organisms (which appear pink to red) from non-lactose fermenters (which appear colorless or normal). Key Features:
Selective:Bile salts and crystal violet inhibit the growth of most Gram-positive bacteria.
Differential:Differentiates lactose fermenting (pink/red) and non-lactose fermenting (colorless) Gram-negative bacteria.
Uses:Isolation and identification of enteric bacteria from various sources like clinical specimens, food, and water.
Principle:Lactose fermentation results in acid production, which lowers the pH and causes the neutral red indicator to turn pink or red in lactose-fermenting bacteria.
Components:Peptone, lactose, bile salts, crystal violet, neutral red, and agar.
How it Works:
Bile salts and crystal violet inhibit the growth of most Gram-positive bacteria, allowing Gram-negative bacteria to grow.
Lactose-fermenting bacteria metabolize lactose, producing acid as a byproduct.
The acid lowers the pH, causing the neutral red indicator to turn pink or red in the colonies of lactose fermenters.
Non-lactose fermenting bacteria do not produce acid, so their colonies remain colorless or the original color of the medium.
Specific Examples:
Escherichia coli is a lactose fermenter, producing pink to red colonies with bile precipitates.
Salmonella and Shigella species are non-lactose fermenters, producing colorless or transparent colonies.
Antibiotic Assay Medium No. 19 is a microbiological medium used for the bioassay of various antibiotics, particularly Amphotericin B, Nystatin, and Candicidin, using Saccharomyces cerevisiae as the indicator organism. It is a nutrient-rich medium providing essential elements for microbial growth and is used in both the base and seed agar for diffusion assays. Here's a more detailed description:Composition:
Peptone (9.4 g/L): Provides a source of nitrogen, vitamins, and amino acids.
Yeast Extract (4.7 g/L): Supplies vitamins, amino acids, and other growth factors.
Beef Extract (2.4 g/L): Contributes to the nutrient base with nitrogen and other essential components.
Dextrose (10.0 g/L): Acts as a carbon and energy source for microbial growth.
Sodium Chloride (10.0 g/L): Maintains the osmotic balance of the medium, crucial for cell integrity.
Agar (23.5 g/L): A solidifying agent that forms a gel base for the assay.
Intended Use:
Antibiotic Assays:Specifically designed for the microbiological assay of antibiotics like Amphotericin B, Nystatin, and Candicidin, which are antifungal agents.
Cylinder Plate Method:Commonly used with the cylinder plate method (also known as the cup-plate method) where antibiotic solutions diffuse from cylinders into the agar, creating zones of inhibition that are measured to determine antibiotic potency.
Seed and Base Agar:Employed as both the base and seed medium in agar diffusion assays.
USP/EP Compliance:Formulated according to the specifications of the United States Pharmacopeia (USP) and the European Pharmacopoeia (EP).
Key Features:
Nutrient-rich:Provides a wide range of nutrients for the growth of Saccharomyces cerevisiae.
Osmotic Balance:Sodium chloride maintains the correct osmotic pressure for cell stability.
Standardized:The formula is adjusted and standardized to ensure consistent and reliable results in antibiotic assays.
BSE/TSE Free:In some formulations, such as Antibiotic Veg Assay Medium No. 19, vegetable peptones replace animal peptones, eliminating risks associated with bovine spongiform encephalopathy (BSE) and transmissible spongiform encephalopathy (TSE).
Pseudomonas Agar for Fluorescein is a specialized growth medium used to identify and cultivate Pseudomonas species, particularly focusing on their ability to produce fluorescein, a fluorescent pigment. It's a selective medium that enhances fluorescein production while inhibiting the formation of other pigments like pyocyanin. Here's a more detailed description:Purpose: To promote the growth and differentiation of Pseudomonas species based on their ability to produce fluorescein, a yellow-green fluorescent pigment. Composition:
Peptones: Provide essential nitrogenous compounds, amino acids, and vitamins.
Dipotassium phosphate: Helps to enhance fluorescein production and suppress pyocyanin.
Agar: Serves as a solidifying agent.
Appearance:
Appearance of the powder: Light beige, free-flowing, and homogeneous.
Appearance of the prepared medium: Light yellow to yellow, slightly opalescent gel.
Mechanism:
The medium is formulated to encourage the production of fluorescein, a water-soluble, chloroform-insoluble pigment produced by some Pseudomonas species.
This pigment diffuses into the agar surrounding the bacterial colonies, creating a characteristic yellow-green fluorescence when observed under ultraviolet light.
Key features:
It is recommended by the U.S. Pharmacopeia for microbial limit tests.
It helps differentiate Pseudomonas species based on their ability to produce fluorescein.
It inhibits the production of pyocyanin, another pigment produced by some Pseudomonas species, allowing for clearer observation of fluorescein production.
MP-5 Medium is a microbiological growth medium used to detect pectinolytic microorganisms, particularly those that produce the enzyme polygalacturonase. It's designed to identify organisms capable of degrading pectin, a component of plant cell walls. This is achieved by observing either a clear zone or halo around the microbial colony, indicating pectin degradation, or by using a precipitant solution that reveals zones of pectin hydrolysis. Here's a more detailed breakdown:
Purpose:MP-5 Medium is specifically formulated to detect pectinolytic activity, which is the ability of microorganisms to break down pectin.
Pectinase Detection:Many bacteria and fungi produce pectinases (enzymes that degrade pectin). MP-5 Medium helps identify these organisms.
Detection Method:The medium contains pectin as a substrate. When pectinolytic microorganisms grow on it, they break down the pectin, resulting in a clear zone or halo around the colony. This can be visualized by adding a precipitant solution (like hexadecyltrimethyl ammonium bromide) that makes the intact pectin opaque, thus highlighting the clear zones where pectin has been hydrolyzed.
Examples:HiMedia Laboratories mentions that certain bacteria like Erwinia carotovora and Erwinia chrysanthemi are known pectin degraders. A document on Scribd indicates that Aspergillus brasiliensis and Fusarium moniliforme are also known to be positive for polygalacturonase production when tested with this medium.
Applications:MP-5 Medium is useful in various applications, including food microbiology, plant pathology, and general microbial identification.
Modified Phosphate Buffer is recommended as washing buffer solution required during detection of Escherichia coli O157 : H7 from food and animal feeding stuffs. The composition and performance criteria are in accordance with ISO 16654: 1999. This buffer solution is used as washing solution for immunomagnetic separations. It is important to keep the pH of the suspending fluid within the same range, as variation in pH may affect the viability of the microorganisms. Phosphate buffer is preferred to unbuffered distilled water in order to standardize wide variation in the pH of distilled water from various sources.
Colourless or white to off-white crystals or powder
Solubility
33.3 mg soluble in 1 mL of water
Potassium persulfate, with the chemical formula K2S2O8, is an inorganic compound also known as potassium peroxydisulfate or KPS. It's a white solid that dissolves in water, and is a strong oxidizing agent and initiator for polymerizations. The Hi-AR/ACS grade indicates a high purity and analytical grade, suitable for various chemical applications. Key Features and Properties:
Chemical Formula: K2S2O8.
Appearance: White solid.
Solubility: Sparingly soluble in cold water, but dissolves better in warm water.
Oxidizing Agent: Potassium persulfate is a strong oxidizing agent.
Polymerization Initiator: Used to initiate polymerization reactions.
Hi-AR/ACS Grade: Indicates high purity and analytical grade, suitable for various chemical applications, says HiMedia.
HybridoXL™ DMSO Sterile Filtered is a sterile, filter-sterilized, hybridoma-tested DMSO used in cell culture for cryopreservation and other applications. It's a cryoprotectant used to prevent ice crystal formation during freezing and to protect cells from damage. DMSO is commonly used in cell freezing media at a concentration of 10% (v/v) and can be used for various cell lines and tissues. Here's a more detailed breakdown:
Function:HybridoXL™ DMSO is a cryoprotectant that prevents ice crystal formation during cell freezing, which can cause damage to cells.
Use:It's used for cryopreservation of animal cells and tissues, including human embryos and blood cells.
Concentration:Typically used at a 10% (v/v) concentration in cell freezing media.
Other Applications:It can also be used to induce cell differentiation and has applications in molecular biology, such as PCR.
Features:HybridoXL™ DMSO is sterile-filtered and hybridoma-tested, ensuring quality and suitability for hybridoma cell line cryopreservation.
Safety:It is generally considered safe for cell culture, with most cell lines tolerating up to 1% DMSO without significant toxicity.
Hanks' Balanced Salt Solution (HBSS) 1X is a buffered salt solution used in various cell culture applications. It maintains pH and osmotic balance, provides essential ions, and can be used for rinsing, diluting, or transporting cells and tissues. HBSS is often used in cell culture to maintain a physiological environment, especially for cells maintained in non-CO2 conditions. Key Features and Uses:
Maintenance of pH and osmotic balance:HBSS helps maintain the proper pH and osmotic pressure within cells, which are crucial for their survival and function.
Provision of essential ions:It provides cells with the necessary inorganic ions, like sodium, potassium, calcium, and magnesium, that are essential for various cellular processes.
Washing, diluting, and transporting cells:HBSS is used to rinse cells before dissociation, dilute cells for counting, and transport cells or tissue samples.
Diluent and reagent preparation:It serves as a diluent for various cell culture reagents and can be used to prepare buffers.
Formulations and Variations:
Phenol Red:Some HBSS formulations contain phenol red, an indicator that changes color with pH changes. Others are phenol red-free.
Calcium and Magnesium:Some HBSS formulations include calcium and magnesium, while others are calcium and magnesium-free.
CO2 dependency:HBSS is designed to equilibrate with air and doesn't require a CO2 environment, making it suitable for cells maintained in non-CO2 conditions.
Common Uses:Washing cells before dissociation or passage, Transporting cells or tissue samples, Diluting cells for counting, Preparing reagents, and Maintaining cells in non-CO2 conditions.
Colourless or white hygroscopic crystals or powder or solid
Form
Solid
Sucrose, also known as table sugar, is a disaccharide made of glucose and fructose. It is widely used in plant culture as a primary energy source, providing carbon and energy for plant cells. Sucrose, particularly for plant tissue culture, is available in different grades (e.g., Grade I, Grade II) and is typically tested for purity and quality. Here's a more detailed look:
Chemical Properties:Sucrose has the chemical formula C12H22O11 and a molecular weight of 342.30.
In Plant Culture:It serves as a major carbohydrate source, providing energy to plant cells during in vitro culture.
Purity:Sucrose used in plant culture is often tested for impurities like heavy metals and sulfate.
Grades:Different grades of sucrose (e.g., Grade I, Grade II) are available, with Grade I typically being of higher purity.
Specific Applications:Sucrose can be used in various plant culture applications, including micropropagation, callus induction, and as a cryoprotectant.
Effects of Concentration:Sucrose concentration in the culture medium can influence plant growth, with optimal concentrations being crucial for various processes like rhizogenesis and callogenesis.
Agar powder is a natural gelling agent derived from red seaweed, primarily used as a plant-based alternative to gelatin. It's a versatile ingredient used in various culinary applications, from thickening liquids to making jellies and desserts, and also plays a crucial role in microbiology as a culture medium. Here's a more detailed look:
Source:Agar is extracted from the cell walls of certain red algae, including Gelidium and Gracilaria.
Composition:It mainly consists of two polysaccharides: agarose and agaropectin, with agarose being responsible for its gelling properties.
Uses in cooking:
Gelling agent: Agar is used to solidify liquids, creating gels and textures in various dishes.
Thickener: It can thicken soups, custards, and other foods.
Vegetarian alternative: Agar is a popular choice for vegan and vegetarian diets as a replacement for gelatin.
Uses in microbiology:
Culture medium: Agar is a fundamental component of culture media used to grow bacteria and other microorganisms in laboratories.
Properties:Agar powder is tasteless, odorless, and colorless, making it suitable for various applications.
Gelation:When dissolved in hot water and cooled, agar forms a stable and firm gel, which melts above 85°C (185°F).
Forms:Agar is available in powder, flake, and bar forms.
How to use:Agar powder dissolves quickly in boiling water and sets into a gel when cooled, making it easy to use in various recipes.
A vented cap on a tissue culture flask facilitates gas exchange, allowing the cells to receive oxygen and release carbon dioxide, which is crucial for their growth and survival. This type of cap differs from non-vented caps, which create a sealed environment to prevent contamination. Vented caps are typically used for cultures that require oxygen and are suitable for short-term experiments or when daily feeding/water exchanges are not needed. Key features of vented caps:
Gas exchange:Allows for the passage of gases, including oxygen and carbon dioxide, between the flask and the surrounding environment.
Sterility:Vented caps often incorporate a filter that prevents microbial contamination while allowing gas exchange.
Types of Vented Caps:
Vent caps with filter membranes: These are common in tissue culture flasks and use a filter membrane (typically 0.22µm) to allow gas exchange while preventing the entry of microorganisms.
Vented caps with small openings: These caps have small, pre-drilled holes or vents that allow for gas exchange but may not offer as robust protection against contamination as filter-equipped caps.
Benefits of Vented Caps:
Adequate oxygenation:Essential for the growth of many cell types, as they require a constant supply of oxygen.
Prevention of buildup of toxic gases:Carbon dioxide buildup can inhibit cell growth, so venting helps maintain a healthy environment.
Simplified handling:Vented caps allow for easier gas exchange during short-term experiments where daily feeding and water changes are not required.
In summary: Vented caps on tissue culture flasks are designed to allow gas exchange while minimizing contamination, making them suitable for various cell culture applications, especially those requiring oxygen and where daily media changes are not needed.
TCL072 is a sterile filtered 50X HAT medium supplement containing 680.5 mg/litre Hypoxanthine, 8.8 mg/litre Aminopterin and 193.8 mg/litre Thymidine in Phosphate Buffered Saline. HAT supplement contains Hypoxanthine, Aminopterin and Thymidine and is used for the preparation of selection medium for hybridoma
DNase I is a deoxyribonuclease enzyme that specifically cleaves phosphodiester bonds in DNA, leading to the degradation of both single- and double-stranded DNA into shorter oligonucleotide fragments. Its activity is heavily influenced by the presence of divalent metal ions, particularly Mg²⁺ and Ca²⁺, which are essential for optimal enzymatic function.Key Features and Properties
Source and Form: DNase I is typically prepared from bovine pancreas and is available either as a lyophilized powder or in solution form (e.g., 1 mg/mL) for immediate use in lab procedures.
Enzymatic Activity: The enzyme has a minimum activity of 3000 U/mg (Kunitz) and releases 5’-phosphorylated and 3’-hydroxylated ends upon digestion 3.
Storage conditions: Generally, it is recommended to store DNase I at -20°C, avoiding frost-free freezers due to temperature fluctuations. Once reconstituted, it should be kept on ice until used and can be aliquoted and frozen for stability 2.
Heat Inactivation: DNase I can be inactivated by heating to 75°C for 10 minutes or by incorporating chelators like EDTA2 .
5 Sources
ApplicationsDNase I plays a critical role in various molecular biology techniques, including:
Removing genomic DNA from RNA preparations prior to reverse transcription or PCR, ensuring downstream applications are DNA-free 1.
Avoiding cell clumping during tissue disaggregation: the enzyme degrades released DNA from ruptured cells, preventing aggregation in concentrated or cryopreserved cell suspensions 2.
Nick Translation and Footprinting Studies: DNase I is utilized in generating marked DNA for labeling purposes and in footprinting assays to study DNA-protein interactions 1.
4 Sources
Handling Instructions
Always use RNase-free and DNase-free materials when working with DNase I to prevent contamination and ensure the integrity of nucleic acids 1.
Avoid vigorous mixing or vortexing of DNase I solutions as it is sensitive to physical denaturation 2.
3 Sources
ConclusionDNase I is an essential reagent in molecular biology for purposes ranging from DNA elimination in RNA protocols to cell dissociation and more. As a versatile tool, its effectiveness is maximized through proper handling and understanding of its properties, ensuring reliable results in experimental applications. For detailed handling instructions and specific protocols, refer to the product datasheets from the respective manufacturers.
Potato Dextrose Agar (PDA) is a microbiological growth medium widely used for cultivating fungi, particularly yeasts and molds. It's a nutrient-rich medium composed of potato infusion, dextrose (glucose), and agar, which serves as a solidifying agent. PDA is favored for its ability to support the growth of various fungi, including those found in food, dairy products, and even clinical samples. Key Components and Purpose:
Potato Infusion: Provides a source of nutrients and growth factors for fungi.
Dextrose (Glucose): Acts as a readily available carbohydrate source for energy.
Agar: A solidifying agent that creates a stable surface for microbial growth.
Applications:
Cultivating Fungi:PDA is a standard medium for growing and isolating fungi, including yeasts and molds.
Enumeration of Fungi:Used to count the number of fungal colonies in a sample.
Studying Fungal Morphology:The medium allows for observation of colony characteristics and pigment production in some fungi.
Fungal Identification:Certain fungal species can be identified based on their growth patterns and colony characteristics on PDA.
Testing for Fungal Contamination:PDA is used in food and dairy industries to assess the presence and level of fungal contamination.
Antimicrobial Agent Testing:Can be used to evaluate the effectiveness of antimicrobial agents against fungi.
Modifications:
Acidification:PDA can be acidified (e.g., with tartaric acid) to inhibit bacterial growth, making it more suitable for fungal isolation.
Supplementation with Antibiotics:Antibiotics like chloramphenicol can be added to suppress bacterial growth, allowing for selective fungal growth.
Copper Supplementation:Some studies have shown that adding copper to PDA can enhance pigmentation in certain fungal cultures, according to a study published in FEMS Microbiology Letters.
A nichrome loop is a laboratory tool, typically made of nichrome wire, used for transferring and manipulating small samples of microorganisms, cells, or other materials. It's commonly used in microbiology for streaking cultures, inoculating media, and other applications. The wire is often formed into a loop shape, and the tool is designed to be easily heated for sterilization. Here's a more detailed description:
Material:Nichrome wire, an alloy of nickel and chromium, is used for its high electrical resistance and ability to withstand high temperatures without oxidizing or breaking.
Shape:The wire is typically formed into a loop, which can be circular or oval, and may have different diameters depending on the intended application.
Handle:Nichrome loops may come with or without handles. Handles are often made of insulated material like aluminum or plastic for easier and safer handling.
Applications:
Microbiology: Transferring microbial cultures to agar plates, inoculating liquid media, and performing flame tests.
Tissue culture: Transferring cells.
Other applications: Nichrome loops can also be used in ultramicrotomy for transferring sections and in some industrial applications like hot-wire foam cutting.
Sterilization:Nichrome loops can be easily sterilized by flaming, making them suitable for repeated use.
Peptone Water, particularly from HiMedia, is a widely used microbiological medium. It serves as a general-purpose growth medium for non-fastidious microorganisms and also acts as a base for carbohydrate fermentation studies. It's composed of peptic digest of animal tissue and sodium chloride, providing essential nutrients like nitrogen and maintaining osmotic balance. Here's a more detailed breakdown:Key Components and Properties:
Peptone: Provides nitrogenous and carbonaceous compounds, amino acids, and vitamins, supporting bacterial growth.
Sodium Chloride: Maintains the osmotic balance of the medium, crucial for cell viability.
pH: Typically adjusted to 7.2 ± 0.2 at 25°C, but can be adjusted for specific applications like Vibrio cultivation (pH 8.4).
Appearance: Usually a light yellow to yellowish-brown powder.
Solubility: Readily soluble in water.
Applications:
General Growth Medium:Supports the growth of a wide variety of microorganisms, especially those not requiring highly specific growth factors.
Carbohydrate Fermentation Studies:A basal medium for studying the ability of organisms to ferment specific carbohydrates. By adding a carbohydrate and a pH indicator (like phenol red), researchers can observe acid production (indicated by a color change) and gas production (using Durham tubes).
Indole Production Testing:Specifically recommended for detecting indole production by Escherichia coli.
Enrichment of Vibrio Species:Peptone water with a pH of 8.4 is particularly useful for cultivating Vibrio species.
Types of Peptone Water:
Peptone Water (general): The basic formulation.
Peptone Water with Phenol Red: Contains phenol red indicator for carbohydrate fermentation studies.
Peptone Water with pH adjusted to 8.4: Specifically for Vibrio species.
Andrade Peptone Water: A basal medium for carbohydrate fermentation with Andrade's indicator.
Buffered Peptone Water: Used for pre-enriching damaged Salmonella species from food samples.
In essence, Peptone Water is a versatile and essential tool in microbiological labs, offering a simple yet effective way to cultivate, identify, and study microorganisms.
Collagenase Type IV is a specialized enzyme that breaks down type IV collagen, a key component of the basement membrane. It's derived from bacteria like Clostridium histolyticum. This enzyme is used in various applications, including isolating islet cells, dissociating tissues, and degrading extracellular matrix. Key Features and Applications:
Specificity:Collagenase Type IV has a specific affinity for the sequence R-Pro-(X-Gly-Pro), where X is often a neutral amino acid, allowing it to target type IV collagen.
Tissue Dissociation:It's effective in breaking down the intercellular matrix, which is crucial for separating cells from tissues.
Basement Membrane Degradation:It can degrade both type IV and type VII collagen, which are major components of the basement membrane.
Applications:
Islet Isolation: Recommended for isolating Islets of Langerhans, which are important for insulin production.
Cell Dissociation: Used in cell culture for isolating cells from tissues, especially when dealing with dense connective tissues.
Stem Cell Research: Used in various stem cell research areas, including reprogramming, maintenance, and differentiation of both human and mouse pluripotent stem cells.
Storage and Preparation:
Powdered Form: Stored at 2-8°C.
Solution: Reconstituted solution should be stored at -20°C.
Stock Solution: A 100mg/mL stock solution can be prepared by dissolving the powder in Hank's Balanced Salt Solution (HBSS).
Filtering: The stock solution should be filter-sterilized before use.
Important Considerations:
Working Concentration:The optimal working concentration needs to be determined based on specific experimental conditions and the type of tissue being dissociated.
Secondary Proteases:Some collagenase preparations may contain clostripain and other secondary proteases, which can be a concern if they damage membrane proteins or receptors. Collagenase Type IV is designed to minimize these activities.
Safety:Collagenase is used in research and may not be suitable for direct human application without proper evaluation and approval.
Casein Soyabean Digest Broth, also known as Soybean Casein Digest Medium (SCDM) or Tryptone Soya Broth (TSB), is a widely used general-purpose microbiological medium for cultivating a broad range of bacteria and fungi. It's a nutrient-rich broth that supports the growth of both fastidious and non-fastidious microorganisms. Key Characteristics and Components:
General-Purpose Medium: SCDM is versatile and suitable for various applications, including sterility testing, microbial limit testing, and general cultivation in research and quality control.
Nutrient Rich: It contains papaic digest of soybean meal and enzymatic digest of casein, providing a rich source of amino acids, peptides, and other nutrients for microbial growth.
Composition: Common ingredients include soybean meal digest, casein digest, dipotassium phosphate, sodium chloride, and glucose.
Appearance: Typically a light beige, free-flowing powder that dissolves in water.
pH: The typical pH of the prepared medium is around 7.3 ± 0.2.
Applications: SCDM is used for:
Cultivating a wide variety of bacteria and fungi.
Sterility testing of pharmaceuticals, food products, and other materials.
Microbial limit testing to determine the total viable count of microorganisms.
General microbiological research and quality control.
Alternative Formats: SCDM is also available in agar form (Soybean Casein Digest Agar or Tryptone Soya Agar) for plate-based cultivation.
Apart from the uses mentioned in section 1.2 no other specific uses are stipulated
Appearance
Cream to yellow coloured homogeneous free flowing powder
pH
7.20 - 7.60
Form / Physical state
Dehydrated powder, cream to yellow, homogeneous, free‑flowing
Standards / Performance
The composition and performance criteria conform to ISO 6880:1983 (as per Hi
Brain Heart Infusion (BHI) Broth is a highly nutritious, buffered liquid culture medium used to grow a wide variety of microorganisms, including fastidious and pathogenic bacteria, fungi, and yeasts. It is made from an infusion of animal heart and brain, providing amino acids, carbon, vitamins, and minerals, along with glucose and a buffering agent like disodium phosphate. BHI Broth is versatile and can be enriched with other components like blood, hemin, or vitamin K to support the growth of specific organisms for diagnostic and research purposes. Key features
Nutrient-rich: Contains the necessary components for a broad range of microbial growth, including amino acids, vitamins, and carbohydrates from brain-heart infusion and glucose.
Buffered: The addition of disodium phosphate helps maintain a stable pH, which is crucial for microbial survival and growth.
Versatile: Can be modified with additives for specific applications.
Common uses
Cultivating microorganisms: Used for the general cultivation of bacteria, including pathogenic cocci and fastidious organisms.
Fungal and yeast growth: Suitable for culturing yeasts and molds; with the addition of sheep blood, it can be used for isolating fungi like Histoplasma capsulatum.
Blood cultures: Used as a medium for setting up blood cultures.
Antimicrobial susceptibility testing: Used to prepare bacterial inocula for susceptibility testing.
Diagnostic tests: Specifically suited for cultivating Staphylococcus for the plasma coagulase test.
Enrichment for specific organisms
For anaerobic growth: Enriched with vitamin K and hemin.
For Haemophilus species: Enriched with Fildes enrichment, hemin, and NAD.
Sterile tissue culture plates are essential tools in laboratory settings for growing cells in vitro. These plates are designed to provide an optimal environment for cell growth and experimentation. Here are detailed descriptions and specifications of some common sterile tissue culture plates:Key Features of Tissue Culture Plates
Material: Most plates are made from high-quality, clear or natural polystyrene, providing excellent transparency for cell observation under a microscope.
Sterilization: All plates are sterilized using gamma irradiation to ensure they are free from contaminants. They are also RNase/DNase free and non-pyrogenic.
Surface Treatment: Plates often feature a treated surface to improve cell adhesion, enhancing growth conditions for adherent cells.
Common Types of Plates
96-Well Plates:
Capacity: Up to 0.39 mL per well.
Specifications:
Wells designed for improved cell adhesion and stability.
Alphanumeric markings on wells for easy identification 2.
Applications: Ideal for high-throughput screening and assays.
24-Well Plates:
Volume: 1.90 mL recommended medium volume per well.
Design: Circular wells with flat bottoms to minimize evaporation and contamination risks 1.
6-Well Plates:
Growth Area: Approximately 9.6 cm² per well, suitable for larger cell cultures.
Features: Raised rims to prevent evaporation and cross-contamination 2.
Round Bottom Plates:
Use: Often used for suspension cell cultures where more mixing is required 1.
5 Sources
Additional Specifications
Well Count: Plates are available in various well counts (6, 12, 24, 48, 96) to accommodate different experimental needs.
Packaging: Typically sold in packs of 50 or 100, individually wrapped to maintain sterility until use 2.
Storage: Plates must be stored in ambient conditions to preserve their integrity 1.
3 Sources
Sterile tissue culture plates are critical for various applications, including cell biology, microbiology, and molecular biology research. When selecting a plate, consider the specific needs of the cell lines and experiments being conducted.
A 70µm cell strainer is a laboratory tool used to separate cells or cell aggregates from a suspension, typically for preparing single-cell suspensions. The 70µm pore size allows passage of single cells and small aggregates while retaining larger debris and cell clumps. This makes it useful for various applications like preparing samples for flow cytometry, isolating specific cell populations, and preparing samples for cell culture or freezing. Key Features and Applications:
Mesh Material: Typically made of strong nylon mesh.
Pore Size: 70µm, allowing passage of most individual cells and small cell aggregates.
Applications:
Flow Cytometry: Preparing single-cell suspensions for analysis.
Cell Culture: Isolating cells for further culturing.
Steam indicator tape, also known as autoclave tape, is a self-adhesive tape used in steam sterilization processes to visually confirm that items have been exposed to the sterilization cycle. It features a chemical indicator that changes color (usually from light yellow to black or dark brown) after exposure to the specific conditions of steam sterilization (temperature and time). This provides a quick and easy way to verify that sterilization has occurred. Here's a more detailed breakdown:Purpose:
Visual Confirmation:The primary function is to provide a clear visual cue that a package or item has been subjected to the steam sterilization process.
Process Indicator:It acts as a process indicator, showing whether the item has passed through the sterilization cycle.
Secure Packaging:In addition to indicating sterilization, it can also be used to securely hold sterilization packs together.
How it works:
Chemical Indicator:The tape contains a chemical indicator ink that reacts to the specific parameters of steam sterilization (temperature and time).
Color Change:Upon exposure to the required conditions, the indicator ink changes color, typically from light yellow to black or dark brown, indicating that the item has been sterilized.
Specificity:The tape is designed to be responsive to steam sterilization and not other sterilization methods.
Key Features:
Adhesive:The tape has a pressure-sensitive adhesive that adheres well to various wrapping materials, including cloth, paper, and some plastics.
Legibility:The tape can often be written on with indelible ink for labeling purposes, and the ink does not smudge.
Versatility:It can be used in various steam sterilizers and with different sterilization cycles.
Examples of Use:
Securing Packs: The tape can be used to seal and hold sterilization packs together.
Identifying Processed Items: It helps differentiate between items that have been sterilized and those that have not.
Confirming Sterilization: It provides a visual confirmation that the sterilization process has been completed.
Important Considerations:
Not a Sterility Assurance:While the tape confirms exposure to steam sterilization conditions, it does not guarantee sterility.
Follow Instructions:Always follow the manufacturer's instructions for the specific tape being used, including appropriate sterilization cycles and storage conditions.
Check for Color Change:Regularly inspect the tape after sterilization to verify the color change and ensure proper exposure.
