Download Filled in by Vector Core: Project: Received: Lot: BIOCENTER

Filled in by Vector Core:
Project:
Received:
Lot:
BIOCENTER KUOPIO NATIONAL VIRUS VECTOR LABORATORY
ADENO-ASSOCIATED VIRUS -VECTOR ORDER FORM
Contact Information
Principal Investigator:
Department/Institution:
Phone/Fax:
E-mail:
Primary Lab Contact:
Department/Institution:
Delivery Address:
Phone/Fax:
E-mail:
Services Requested
a. Vector name:
b. Expression cassette (promoter+transgene):
c. Requested vector preparation quantity (one concentrated prep = 100 µl, titer *1011-1012 vg/ml)
___
Qty of concentrated preps
d. Titer determination
___
FACS (vectors with fluorescent marker gene)
___
Q-PCR
e. Vector testing
___
Sterility
___
Replication competent virus
Biosafety and Compliance
___ I have read and approved Virus Vector MTA (attach a signed copy of MTA).
To comply with guidelines and to be aware of the likely hazards in case of incidental exposure (e.g. to lab personnel
or environment) vector requester is responsible of performing risk assessment of the genes of interest included in
vectors following national guidelines (www.geenitekniikanlautakunta.fi). Please attach a completed copy of BCK
Vector Insert Biosafety Form.
____ BCK Vector Insert Biosafety Form attached.
The materials produced by the BCK Virus Vector Laboratory require the following levels of containment:
BSL1: plasmids and virus vectors tested free of replication competent virus (upon specific request)
BSL2: virus vectors upon standard request
if the inserted genetic material does not increase the risk.
Does the vector insert pose any special handling considerations (e.g. oncogenic or toxic transgene product)?
____ Yes
____ No
Is the biosafety level of the viral vector influenced by the nature of the inserted genetic material?
____Yes
____No
The requester is responsible to have an approval to possess and work with genetically modified organisms according
to national guidelines.
I have been informed that the materials requested may be biohazardous and I have consulted the guidelines.
Signature of Requester:
Plasmid vector for virus production
Plasmid vector backbone used for cloning of the insert:
____
pAAV-CMV
____
other
Please specify (attach vector map):__________________________________
Size of the insert (bp): _______________
Size of the plasmid (bp): ______________
Please submit at least 1 mg of vector plasmid (conc. > 1 µg/µl) per requested virus preparation quantity. Accepted
plasmid purification methods are Qiagen Endo-free Maxi/Mega/Giga protocols, CsCl purification or equivalent
endotoxin-free protocol. Plasmid DNA should be analyzed by restriction enzyme digestion and checked for purity
(A260/280 > 1.9).
Plasmid name and lot:
Method of purification:
Plasmid concentration (by OD260):
A260/280 ratio:
Volume delivered:
Attach gel photo with plasmid:
1. Undigested
2. Linearized
3. Digested with appropriate REs to confirm correct insert size (please specify REs and provide a plasmid
map).
____ Plasmid propagation and purification is requested to be done by BCK Virus Vector Laboratory (price available
by request).
Please attach an aliquot of vector plasmid, plasmid sequence and map indicating restriction enzymes
feasible for digestion analysis.
Plasmid name:
Concentration and volume:
E.coli strain used in propagation:
Titering of vectors
Titering of the AAV-vectors is done with Q-PCR using CMV-primers and SYBR-green. If your construct has an
alternative promoter you will need to provide optimized PCR primers and Q-PCR protocol for your construct.
Please contact Vector Laboratory for detailed instructions.
For any questions or concerns contact the Virus Vector Laboratory.
VIRUS VECTOR LAB CONTACT INFO
Adeno:
Johanna Laakkonen, PhD
[email protected]
Lenti:
Petri Mäkinen, PhD
[email protected]
AAV:
Tommi Heikura, PhD
[email protected]