Agrobacterium rhizogenes Liquid Medium - BioReagent

Cat. No.: A1528482

GRADE & PURITY BioReagent

Storage

Room temperature

Shipped In

Normal

Panoramica

For the cultivation of Agrobacterium rhizogenes

Specifications

Specifiche e purezza

BioReagent

Condizioni di conservazione di stoccaggio

Room temperature

Spedito in

Normal

Grado

BioReagent

Product Line

Prodotti biologici

Application Protocols

The following are general, literature-based protocols for using a liquid Agrobacterium medium. They are provided as guidance and are not product specifications. Adjust to your strain and workflow.

A. Starter culture and expansion

1. Inoculate 5–10 mL of the liquid medium with a single A. rhizogenes colony or 50–100 µL of glycerol stock.
2. Incubate at 26–28 °C, 180–220 rpm, 12–18 h until OD600 ~0.6–1.2 (mid- to late-log; calibrate for your system).
3. Expand to 50–200 mL in baffled flasks at ≤20% fill volume; continue shaking to desired OD.

B. Optional virulence induction

1. Add acetosyringone from a sterile-filtered stock to 50–200 µM final concentration.
2. Incubate 1–3 h before co-cultivation with plant tissue.

C. Antibiotic selection (if applicable)

• Add appropriate antibiotics for plasmid maintenance at validated concentrations for Agrobacterium. Composition and baseline resistance vary by strain; verify MIC.

D. Preparation for co-cultivation

1. Harvest cells by gentle centrifugation (e.g., 3,000–5,000 × g, 10 min), resuspend in induction buffer or fresh medium to target OD.
2. Proceed with plant explant infection per your transformation protocol.

Notes

• Sterility, exact medium composition, and recommended additives are not specified for this item; consult CoA and validate in small-scale trials.

Biological Roles

Applicability

• The product is a growth medium; it does not possess a single biological “role” as a molecule. Its function is to support the biology of Agrobacterium rhizogenes.

General biology of Agrobacterium rhizogenes (literature)

• A. rhizogenes is a soil bacterium capable of transferring T-DNA from its Ri (root-inducing) plasmid into plant cells, resulting in hairy root formation. This natural gene transfer system underpins numerous plant biotechnology applications.
• Growth requirements: Like many Agrobacterium species, it grows aerobically at ~22–28 °C in nutrient media containing a suitable carbon source, nitrogen, minerals, and vitamins. Specific growth rates and doubling times depend on strain and medium composition.
• Virulence regulation: Phenolic compounds from wounded plant tissues (e.g., acetosyringone) can induce vir gene expression, enhancing T-DNA transfer efficiency during co-cultivation.

How the medium supports these roles (general)

• The medium provides balanced nutrients to achieve robust log-phase growth, yielding reproducible cell densities for transformation workflows.
• Buffering and ionic balance (if present) stabilize pH during growth and may influence virulence induction efficiency. Exact buffering system and pH for this item are not specified; verify on the CoA if critical for your protocol.

Buffer Applications

Applicability

• This product is a complex nutrient medium, not a defined buffering reagent.

Guidance (general)

• While many media incorporate buffering capacity to maintain near-neutral pH during bacterial growth, the specific buffer identity, pH setpoint, and capacity for this item are not specified; refer to CoA/Spec Sheet.
• For analytical or biochemical buffering needs (e.g., enzyme assays, electrophoresis), use defined buffers (e.g., phosphate, HEPES, MOPS) at known concentrations and pH; do not substitute nutrient media.
• If pH control during Agrobacterium growth is critical, independently verify pH before and after incubation and, if necessary, supplement with compatible buffer components validated for microbial culture.

Green Alternatives

Context

• Environmental considerations for culture media relate to packaging, sterilization energy, waste handling, and shipping weight rather than solvent hazards.

Comparison of options (general/literature)

• Liquid ready-to-use media
  • Advantages: Time savings; consistent lot-to-lot hydration; reduced risk of preparation errors; no autoclave cycle needed if supplied sterile (sterility not specified for this item).
  • Trade-offs: Higher shipping mass/volume; more plastic if supplied in single-use bottles; limited shelf-life once opened; energy embedded in transport.
• Dehydrated (powdered) media
  • Advantages: Lower shipping footprint; longer shelf stability; user controls water quality and sterilization; reduced storage volume.
  • Trade-offs: Requires energy/water for dissolution and sterilization; potential for preparation variability and operator exposure to dust.

Greener practices

• Consolidate shipments and order appropriate volumes to reduce transport emissions.
• If sterility must be assured, prefer membrane filtration over repeated autoclave cycles to save energy where validated.
• Use recyclable packaging and participate in container take-back programs when available.
• Autoclave biological waste efficiently (full loads, validated cycles) and segregate to minimize volume.

Item-specific environmental data: Not specified for this item; consult SDS or sustainability documentation if required.

Pharmaceutical Uses

Applicability

• Not intended for pharmaceutical formulation or as an excipient. For research use only (per Product Data).

General note

• Culture media are occasionally used in pharmaceutical microbiology for environmental monitoring and QC testing, but such uses require media with specified pharmacopeial compliance (e.g., USP/EP) and documented sterility, growth-promotion testing, and composition. These attributes are not specified for this item; users operating under GMP should source media with explicit pharmacopeial status and validation documentation.

Recommendation

• If your application involves regulated manufacturing or release testing, request detailed quality documentation and consider media formulated and tested to compendial standards.

Physical Properties

Item-specific facts (from Product Data)

• Physical state: Liquid medium (aqueous). Appearance details not specified.
• pH, osmolality, conductivity, refractive index, density: Not specified for this item; refer to CoA/Spec Sheet.

General properties (literature/typical for similar Agrobacterium media; not product specifications)

• Matrix: Water-based, typically near-neutral pH (commonly ~6.8–7.2 in many bacterial media; actual value depends on formulation and manufacturer control).
• Density: Approximately that of water (~1.0 g/mL at 20–25 °C) for dilute nutrient solutions.
• Solubility/miscibility: Fully miscible with water; miscibility with most polar protic solvents is high but use of organic solvents is not recommended for culture integrity.
• Thermal behavior: Intended for use at ambient to 28–30 °C culture temperatures. Avoid freezing unless manufacturer indicates compatibility; repeated freeze–thaw can precipitate components.

Practical notes

• Do not treat literature values as specifications. If downstream applications (e.g., OD600 calibration, viscosity-sensitive bioprocess steps) require exact physicochemical parameters, verify with the CoA or measure on receipt under your conditions.

Quality and Grades

Item-specific facts (from Product Data)

• Grade: BioReagent.
• Sterility, endotoxin, bioburden, and exact composition: Not specified for this item; refer to CoA/Spec Sheet.

What BioReagent grade means (general guidance)

• BioReagent designation typically indicates suitability for common biological laboratory procedures, with attention to raw material selection and processing that avoid contaminants detrimental to cell or microbial work. It does not inherently guarantee sterility, cell culture–tested performance, or pharmacopoeial compliance unless explicitly stated.

How to use this information

• Application fit: Appropriate for routine microbiology and molecular biology workflows involving Agrobacterium rhizogenes, including culture expansion and preparations for plant transformation experiments.
• When to request more data: If your workflow requires defined composition, low bioburden, or absence of specific inhibitors (e.g., heavy metals, preservatives), request the lot CoA/Spec Sheet. For regulated work (GMP/GLP), seek documentation on quality system, traceability, and test methods.
• Stabilizers/preservatives: Not specified for this item; confirm presence/absence on the CoA to avoid interference with downstream assays (e.g., antibiotic sensitivity, virulence induction).
• UV background/fluorescence: Not typically specified for media; if optical assays are planned (e.g., OD or fluorescence reporters), validate background in your instrument with your lot.

Reaction and Applications

Applicability

• This section is repurposed to focus on biological applications, as the item is a culture medium rather than a chemical reagent for synthesis.

Typical applications with Agrobacterium rhizogenes (literature/general)

• Bacterial cultivation: Expansion of A. rhizogenes for downstream plant transformation workflows, including induction of hairy roots via Ri plasmid T-DNA transfer.
• Pre-induction: Growth of cultures to mid-log phase and optional virulence induction with phenolic compounds (e.g., acetosyringone) prior to co-cultivation with plant explants.
• Plasmid maintenance: Selection of engineered strains carrying binary vectors using user-added antibiotics (e.g., kanamycin, spectinomycin), with appropriate concentrations validated for the strain and medium.
• Glycerol stocks: Preparation of cryostocks (e.g., 15–25% glycerol, −80 °C) from cultures grown in the medium, for long-term strain archiving.

Practical tips (general)

• Inoculum quality: Start from a single colony or a well-characterized glycerol stock to ensure genetic uniformity.
• Growth phase control: Monitor OD600 to target mid-log (commonly ~0.4–1.0) before induction or infection; stationary-phase cultures often reduce transformation efficiency.
• Asepsis: Use sterile technique to preserve medium quality and experimental reproducibility.

Item-specific composition, antibiotics, and inducers: Not specified for this item; user adds as needed.

Reaction Conditions

Applicability

• Interpreted here as culture/growth conditions for Agrobacterium rhizogenes rather than chemical reaction conditions.

Typical culture conditions (literature/general guidance; not product specifications)

• Temperature: 25–28 °C for routine growth; some strains grow from ~22–30 °C.
• Aeration: Shaking incubator at 180–220 rpm for baffled flasks filled to ≤20% working volume to ensure oxygen transfer; or sparged/impeller-mixed bioreactors maintaining dissolved oxygen >20%.
• Inoculum: Start from a single colony into 3–10 mL medium; expand to larger volumes once exponential growth is observed.
• Monitoring: Measure OD600; mid-log phase commonly at OD600 ~0.4–1.0 depending on pathlength and instrument. Calibrate OD to CFU/mL for your setup if needed.
• Induction (optional): Add acetosyringone to achieve 50–200 µM final concentration during late log phase or immediately before co-cultivation to enhance T-DNA transfer (optimum varies by strain and plant species).
• Antibiotic selection (if using binary vectors): Add appropriate antibiotics at validated concentrations for Agrobacterium (commonly lower than for E. coli). Verify compatibility with the medium.

Item-specific parameters (pH, supplements, sterility): Not specified for this item; confirm on the CoA/Spec Sheet.

Safety and Handling

Item-specific facts (from Product Data)

• GHS/Signal word/H-statements/Pictograms: Not specified for this item; refer to SDS.
• Storage: Room temperature. Shipped under normal conditions.
• Research use: For research use only.

General safety guidance (literature/standard lab practice)

• Biosafety: Treat as a non-sterile or potentially contaminable microbiological medium. Use aseptic technique to avoid unintended microbial growth. Work in a biosafety cabinet when handling live Agrobacterium cultures as per institutional BSL-1/BSL-2 policies.
• PPE: Lab coat, gloves, and safety glasses. Avoid ingestion and inhalation of aerosols. Wash hands after handling.
• Incompatibilities: Avoid mixing with strong oxidizers or acids/bases that can degrade organic components. Do not add sodium hypochlorite directly to nutrient-rich liquids; chlorine gas can evolve if acid is present.
• Sterilization and decontamination: Surfaces—70% ethanol or appropriate disinfectant. Liquid waste—autoclave (e.g., 121 °C, ≥20 min) before disposal per local regulations. If medium contains antibiotics (user-added), dispose as hazardous bio/chemical waste per policy.
• First aid overview: Skin/eye contact—rinse with water. Inhalation—move to fresh air. Ingestion—rinse mouth. Seek medical attention if symptoms persist. Always consult the SDS for authoritative hazard and response information.
• Special risks: Warm, nutrient media readily support microbial proliferation. Clearly label containers and minimize storage time after opening to reduce bioburden.

Solvent Selection

Applicability

• This product is a ready-to-use aqueous liquid medium, not a solvent. Conventional solvent selection does not apply.

Useful context for medium handling (general/literature)

• Polarity: Strongly polar, water-based; fully miscible with water. Not compatible with most organic solvents from a biological perspective, as they can inhibit bacterial growth.
• Additives: If adding sterile stock solutions (e.g., antibiotics, acetosyringone), use water or sterile-filtered, biocompatible cosolvents at low percentages. Confirm compatibility; many Agrobacterium protocols add acetosyringone from a DMSO or ethanol stock to a final concentration commonly 50–200 µM (literature), keeping solvent <1% v/v to avoid growth inhibition.
• Filtration: If in-house sterile filtration is required, use 0.22 µm PES or PVDF units validated for aqueous solutions. Pre-wet membranes per manufacturer guidance.

Comparison (general)

• Liquid vs. dehydrated media: Liquid media are convenient and consistent but heavier to ship/store; dehydrated powders offer longer shelf-life and flexibility but require accurate dissolution and sterilization.

Storage and Reconstitution

Item-specific facts (from Product Data)

• Storage conditions: Room temperature.
• Shipped in: Normal conditions.
• Reconstitution: Not applicable—this is a liquid medium.
• Additional storage details (shelf-life, sterility after opening, preservatives): Not specified for this item; refer to CoA/Spec Sheet.

General guidance for liquid media (literature/practice)

• Upon receipt: Inspect container integrity. If sterility is required, aseptically dispense into sterile vessels or sterile-filter through a 0.22 µm filter before use (only if compatible and needed).
• After opening: Minimize ambient exposure; work aseptically. If allowable by the product, store tightly closed at 2–8 °C to slow microbial contamination, then equilibrate to use temperature before inoculation. Note: This item’s labeled storage is room temperature; refrigeration after opening is a common laboratory practice but confirm compatibility with the manufacturer guidance.
• Light sensitivity: Some vitamins and indicators are light-sensitive; store in the original amber or opaque container when applicable.
• Freeze–thaw: Avoid freezing unless the CoA indicates tolerance; precipitation can occur and may affect performance.
• Aliquoting: For repeated use, aliquot into sterile containers to reduce contamination risk and cumulative exposure.

Always defer to the lot-specific CoA/Spec Sheet for definitive storage and handling instructions.

Structure and Identity

Item-specific facts (from Product Data)

• Product type: Liquid culture medium formulated for Agrobacterium rhizogenes growth and handling (mixture; not a single defined chemical).
• Grade: BioReagent.
• Storage: Room temperature. Shipped under normal conditions.
• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula/MW/SMILES/InChIKey: Not applicable to complex media; not specified for this item.

General description (literature/typical for this class)

• Composition nature: Aqueous solution containing carbon, nitrogen, salts, vitamins, and buffer components suitable for fastidious growth of Agrobacterium species.
• Structural features (conceptual): Multicomponent water-based solution; contains multiple ionic species (e.g., phosphate, ammonium, magnesium) and organic nutrients (e.g., peptone/yeast extract/carbohydrate) depending on formulation. No discrete small-molecule “structure” applies.
• Format: Ready-to-use liquid medium intended for direct inoculation; supplements (e.g., antibiotics, acetosyringone) are user-added as needed for selection or virulence induction. (Supplements not included unless explicitly stated; not specified for this item.)

Notes

• Being a complex mixture, conventional structure identifiers (CAS, CID, InChIKey, SMILES) do not apply. Always consult the lot-specific Specification/CoA for exact composition if required by your quality system.

Synthetic Utility

Applicability

• Not applicable to chemical synthesis. This product is a biological growth medium and is not employed as a reagent or building block in organic or inorganic synthetic chemistry.

Practical redirection

• For content relevant to this product, refer to the sections on Reaction & Applications (biological context), Application Protocols, and Storage & Reconstitution.

Target Specificity

Applicability

• Not an antibody, oligonucleotide, or affinity reagent. Target specificity does not apply to this culture medium.

Item-specific note

• No antigen/epitope, clone, isotype, or species reactivity information is relevant to this product type.