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Construction and Packaging Service of virus Vector

Construction and Packaging Services of AAV

Large scale AAV production

Biosafety
Recombinant viruses are replication deficient and can infect a broad range of mammalian cell types. Please refer to your institutions Occuptation Health and Safety office for guidance on safe handling of viral particles. BrainVTA sends all viral particles on dry ice, If your virus did not arrive on dry ice, please contact us immediately.

Safety
 Adeno-Associated Virus (AAV)
Recombinant AAV constructs do not encode for either a potentially tumorigenic gene product or a toxin molecule. According to guidelines from the National Institutes of Health (NIH), recombinant AAV vectors can be handled in a Biosafety Level 1 (BSL-1). If dealing with biohazardous material then please handle under Biosafety Level 2 (BSL-2) containment.
 Lentivirus (LV)
Lentiviral vector constructs are derived from HIV and are therefore highly efficient vehicles for in vivo gene delivery. They can integrate transgenes into dividing and non-dividing cells, so work with lentiviral vectors must be carefully. NIH guidelines require the maintenance of a BSL-2 or BSL-2 enhanced control facility for work involving lentivirus.

Storage
● For short-term storage, all vectors could be stored at room temperature or 4°C. 
● For long-term storage, keep all vectors at -80°C. Do NOT store at -20°C. Research suggests that long-term storage at -20°C may result in decreased transduction efficiency of the virus.
We recommend aliquoting your virus into desired volumes, snap-freezing in liquid nitrogen or a dry ice/ethanol bath before storing at -80°C for use in future experiments. Please avoid unnecessary freeze-thawing of the vectors, as excessive freeze-thawing cycles may result in a significant decrease in titer and loss of biological activity.

Handling
● Thawing prior to use
On ice or at room temperature, and use immediately.
● During experiment
Keeping on ice at all times.
● Cell Transduction
AAV particles from both small scale and large scale packaging can be applied directly for cell transduction. 
Relative in vitro infectivity1 of AAV vectors:
Cell Line AAV-1    AAV-2 AAV-3 AAV-4 AAV-5 AAV-6 AAV-8 AAV-9 AAV-DJ AAV-DJ/8
Huh-7 13 100 2.5 0.0 0.1 10 0.7 0.0 500 0.2
HEK293 25 100 2.5 0.1 0.1 5 0.7 0.1 500 0.3
HeLa 3 100 2.0 0.1 6.7 1 0.2 0.1 667 0.2
HepG2 3 100 16.7 0.3 1.7 5 0.3 ND 1250 0.5
Hep1A 20 100 0.2 1.0 0.1 1 0.2 0.0 400 0.1
911 17 100 11 0.2 0.1 17 0.1 ND 500 0.0
CHO 100 100 14 1.4 333 50 10 1.0 25000 5.0
COS 33 100 33 3.3 5.0 14 2.0 0.5 500 0.3
MeWo 10 100 20 0.3 6.7 10 1.0 0.2 2857 1.0
NIH3T3 10 100 2.9 2.9 0.3 10 0.3 ND 500 0.1
A549 14 100 20 ND 0.5 10 0.5 0.1 1000 0.1
HT1180 20 100 10 0.1 0.3 33 0.5 0.1 333 0.2
Monocytes 1111 100 ND ND 125 1429 ND ND 100 ND
Immature DC 2500 100 ND ND 222 2857 ND ND 200 ND
Mature DC 2222 100 ND ND 333 3333 ND ND 100 ND
Animal Injections
Provided virus is provided ready for injection in biocompatible F68/PBS. The recommended titer of AAV for in animal injection is 1012 with 300 nL.

50% discount for Pre-Made AAVs