Product Manuals & MSDS

User manual for ready-to-use products and material safety data sheets.

AAV Product Manual

Contents and Storage

AAV stocks are supplied in liquid form at indicated titer. The storage solution PBS / 5% glycerol. Store at - 80°C. If desired, aliquot viral stock upon arrival, and store those aliquots at -80°C freezer immediately.
DO NOT FREEZE AND THAW REPEATEDLY. 

AAV GC particles to be used = MOI (multiplicity of infection) * # of cells to be infected 

Example: If you intend to infect 1 million cells using MOI of 1,000 , you need 1,000 x 1,000,000 = 109 GC for the infection. If the original stock is 1013 GC/ml , then you will need 0.1 ul of the original stock for the dilution. 

24-well plate: 0.2-0.3 ml 

12-well plate: 0.5-0.8 ml

6-well plate: 1-1.5 ml/well 

60mm-plate: 3-4 ml/plate 

Incubate cells with the virus-containing media for 6-12 hours, or as long as you wish. (Optional), you could remove virus-containing media and replace it with fresh, desired media. 

The appropriate amount of viruses used for infecting cells is critical for the outcome of your experiments. The goal is to get 100% of infection without causing any undesired effects. The optimal concentration differs dramatically between cell types for different serotype of AAV. A range of 2,000-50,000 MOI is used for most cell lines for AAV2, but up to 500,000 MOI may be used for some cells. 

To determine this optimal concentration of virus for your study, you could conduct pilot testing in your cell line by using reporter AAV like AAV-GFP. 

Important Notes

AAV stock can be added directly to cells in culture medium (in the presence or absence of serum). 
It is not necessary to remove viruses, change or add medium following infection, although viruses can be removed after 6-12 hours post infections. 
It can take 3-7 days after the AAV infection to detect the gene over-expression

AAV Material Safety Data Sheets

Emergency Contact
484-325-5100

Safety Information

Cultures of replication defective AAV vectors are non-infectious and are not hazardous materials as defined by OSHA 1919.1200. However, these materials are produced in cells where there is the possibility of recombination to form wild type virus. As such, they should be handled as potentially infectious material. 

AAV vectors consist of recombinant transgene sequences (e.g., marker or human genes) flanked by the AAV inverted terminal repeats. The removal of the majority of viral structural genes renders the vector replication-defective and dependent on an AAV helper virus. AAV cultures are normally provided as purified viral particles in phosphate buffered saline at a concentration of up to 1×1013-14 GC/ml. The viral stock consists of particles containing the vector genome (full capsids) and a variable number of empty viral capsids in PBS. Other trace components present include, but are not limited to, inorganic salts, vitamins and other nutrients, and human cellular proteins, carbohydrates, amino acids, and fats. The material is normally shipped and stored frozen. 

None

Liquid or frozen particle suspensions

AAV cultures are non-pathogenic, and are not known to cause any diseases in humans or animals.

None

Stable. Will enter mammalian cells in the presence of adenovirus and wild type AAV can integrate into host cell DNA.

Spill: Contain spill and decontaminate the area using a disinfectant such as chlorine bleach (10% f.c.), Wescodyne, or detergent-based disinfectant.

Waste Disposal: Dispose of viral stock by autoclaving at 121°c for 30-45 minutes Dispose of infected liquid cultures by decontamination with chlorine bleach (10% f.c.) for 10 minutes and then dispose of in sink.

Dispose of infected animal carcasses or tissues by incineration Follow all Federal, State, and Local regulations.
Recombinant AAV (all serotypes) constructs, in which the transgene does not encode either a potentially tumorigenic gene product or a toxin molecule and are produced in the absence of a helper virus can be handled in Biosafety Level 1 facility. Otherwise it should be handled as biohazardous material under Biosafety Level 2 containment.

Vectot BioLabs recommends that all AAV vectors and cultures be handled by qualified biologists using appropriate safety procedures and precautions. Detailed discussions of laboratory safety procedures are provided in Laboratory Safety: Principles and Practice (Fleming et al., ASM Press, 2009), and in the U.S. Government Publication, Biosafety in Microbiological and Biomedical Laboratories (CDC, 2020). This and other publications are available at the Centers for Disease Control Office of Health and Safety’s website at

https://www.cdc.gov/labs/pdf/SF__19_308133-A_BMBL6_00-BOOK-WEB-final-3.pdf

Information on the classification of human etiologic agents on the basis of hazard can be found as Appendix Bin the NIH Guidelines for Research Involving Recombinant DNA Molecules at

https://osp.od.nih.gov/wp-content/uploads/NIH_Guidelines.pdf

The above information is accurate to the best of our knowledge. All materials and mixtures may present unknown hazards and should be used with caution. The user should exercise independent judgment as to the hazards based on all sources of information available. Vector BioLabs shall not be held liable for any damage resulting from the handling or use of the above product.

Adenovirus Product Manual

Contents and Storage

Recombinant adenovirus is supplied in liquid form at indicated titer. The storage solution is DMEM/2.5% glycerol. Store at -80°C. If desired, aliquot viral stock upon arrival, and store those aliquots at -80°C freezer immediately. 
DO NOT FREEZE AND THAW REPEATEDLY. 

PFU needed = MOI (multiplicity of infection) * # of cells to be infected e.g. If you intend to infect 1 million cells using MOI of 100 , you need 100 x 1,000,000 = 108 PFU for the infection. If the original stock is 1010 PFU/ml , then you will need 10 ul of the original stock for the dilution.

24-well plate: 0.2-0.3 ml 

12-well plate: 0.5-0.8 ml

6-well plate: 1-1.5 ml/well 

60mm-plate: 3-4 ml/plate 

60cm-plate: 8-12 ml/plate

Incubate cells with the virus-containing media for 6-12 hours, or as long as you wish. (Optional), you could remove virus-containing media and replace it with fresh, desired media. 

The appropriate amount of viruses used for infecting cells is critical for the outcome of your experiments. The goal is to get 100% of infection without causing cytotoxicity or other undesired effects. The amount of adenovirus cell surface receptors vary greatly among different cell types therefore the optimal concentration differs dramatically between cell types. A range of 20-200 MOI (multiplicity of infection) is used for most cell lines, but up to 4000 MOI may be used for some cell lines or primary cells. 

To determine this optimal concentration of virus for your study, you could conduct pilot testing in your cell line by using reporter adenoviruses, such as GFP adenovirus (Cat.#1060). 

Important Notes

Recombinant adenovirus is for delivering interested genes into mammalian cells. It provides the following advantages:

  1. 100% efficiency of gene delivery in many cell types.
  2. Recombinant viruses can be added directly to cells in culture medium (in the presence or absence of serum).
  3. It is not necessary to remove viruses, change or add medium following infection, although viruses can be removed after 6-12 hours post infections.   

AAV Material Safety Data Sheets

Emergency Contact
484-325-5100, or 877-BIO-LABS

Product Identification:
All pre-made & custom made Ad5 adenovirus by Vector BioLabs.

Biological Name: Adenovirus – Type 5

Characteristics:
Adenoviridae; non-enveloped, icosahedral virions, 75–80 nm diameter, double-stranded, linear DNA genome. The recombinant viruses are based on human adenoviral backbone which is deleted in the essential E1 gene as well as the E3 gene. The viruses produced are thus non-replicative.

Pathogenicity:
Varies in clinical manifestation and severity; symptoms include fever, rhinitis, pharyngitis, cough and conjunctivitis. The risk from infection by defective recombinant adenoviral vectors depends both on the dose of virus and on the nature of the transgene.
Adenovirus does not integrate into the host cell genome but can produce a strong immune response.

Host range:
Humans and animals

Incubation period:
From 1–10 days

Mode of transmission:
In the laboratory, care must be taken to avoid spread of infectious material by aerosol, direct contact or accidental injection

Chemical listed as carcinogen or potential carcinogen:
None

Drug susceptibility:
No specific antiviral available

Susceptibility to disinfectants:
Susceptible to 1% sodium hypochlorite, 2% glutaraldehyde. Recommend use of 1/3 volume of bleach for 30 minutes.

Physical inactivation:
Sensitive to heat; 1 hour at 56°C is used to inactivate virus.

Survival outside host:
Adenovirus type 5 survived from 3–8 weeks on environmental surfaces at room temperature.

Surveillance:
Monitor for symptoms; confirm by serological analysis


First Aid/Treatment

Contact: Immediately flush eyes and skin with plenty of water for at least 15 minutes. Call a physician.

Inhalation: N/A

Ingestion: Wash out mouth with water. Call a physician

Accidental injection: Wash area with soap and water. Call a physician.

Pour 1 volume of Javel water over the leak(s) and wait for 15 minutes. Wipe up carefully.

Hold for autoclave waste disposal and decontaminate work surfaces with 70% alcohol.

Containment requirements:
Biosafety level 2 practices and containment facilities for all activities involving the virus and potentially infectious body fluids or tissues. This level consists of etiological agents considered to be of ordinary potential harm.

Protective clothing:
Recombinant adenovirus: Laboratory coat; gloves.

Other precautions:
Access to the laboratory is limited.
Work surfaces are decontaminated before and after each procedure
Mechanical pipetting devices are used for all procedures; mouth pipetting is prohibited.
Eating, drinking, and smoking are not permitted in the laboratory; food is not stored in laboratory areas.
Laboratory coats are worn in and are removed before leaving the laboratory. Hands are washed before and after handling virus.

Disposal:
Decontaminate all wastes before disposal; steam sterilization

Storage:
In sealed containers that are appropriately labeled

The above information and recommendations are believed to be accurate and represent the most complete information currently available to us. All materials and components may present unknown hazards and should be used with caution. Vector BioLabs, Inc assumes no liability resulting from use of the above products.