User Manual: Ready-To-Use Recombinant Adenovirus
Contents
1. 2011 SignaGen Laboratories SignaGen Laboratories F 9601 Medical Center Drive Recombinant Adenovirus eter ae Storage Instructions and Infection Protocol Rockville MD 20850 FAX 301 560 4919 TEL 301 330 5966 Toll Free 1 866 918 6812 Email info signagen com User Manual Ready To Use 2 2 Web www signagen com This product is for laboratory research ONLY and not for diagnostic use 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 Description 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 In vitro Infection Protocol 1 Prepare virus containing media Thaw viral stock at either room temperature or on ice Add desired amount of virus to media If needed viruses could be diluted further in DMEM or other media 2 Infecting cells with adenovirus Remove the original cell cultu2. re media and add the above virus containing media to cell culture Below isa general guideline for the amount of media used 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 10cm 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 10 100 MOI multiplicity of infection is used for most cell lines but up to 1000 MOI may be used for lymphoid cell lines 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 Ad CMV GFP Cat SL100708
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