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FILLED BY VECTOR CORE PROJECT: RECEIVED: LOT: BIOCENTER KUOPIO NATIONAL VIRUS VECTOR LABORATORY LENTIVIRAL VECTOR ORDER FORM Contact Information Principal Investigator: Department/Institution: Primary Lab Contact: Department/Institution: Delivery Address: Phone/Fax: E-mail: 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 >5x108 TU/ml) ___ Qty of concentrated preps d. Titer determination ___ FACS (vectors with fluorescent marker gene) ___ p24 ELISA e. Vector testing ___ Sterility ___ Replication competent virus (p24 ELISA based infectivity assay, additional price 1500 €) 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 envinronment) 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: ____ LV-PGK ____ other 3rd generation lentiviral vector plasmid Please specify (attach vector map):__________________________________ Size of the insert (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: 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]