User Manual for Ready-To-Use Recombinant
Contents
1. VECTOR BIOLABS THE ADENOVIRUS COMPANY User Manual for Ready To Use Recombinant Adenovirus 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 IMPORTANT GUIDELINES Follow these guidelines when performing infections 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 virus Remove the original cell culture media and add the above virus containing media to cell culture Below is a 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 c2. ontaining 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 tyoes 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 B gal adenovirus Cat 1080 or GFP adenovirus Cat 1060 Date of revision Jan 30 2005 3701 Market Street Suite 340 Philadelphia PA 19104 Tel 877 biolabs Fax 215 525 1112 info vectorbiolabs com
Download Pdf Manuals
Related Search