Takara Adeno-X Adenoviral System 3 (Universal) \10 Rxns

Adeno-X Adenoviral System 3 (Universal) provides the components necessary to place an expression cassette of your choice in an adenoviral vector format. The system provides a perfect platform for the tissue-specific expression of your gene of interest, or for the efficient delivery of shRNA or microRNA. pAdenoX-PRLS (included in the vector set) is a prelinearized adenoviral vector that is ready for the insertion of your expression cassette via our In-Fusion HD PCR Cloning technology. Simply PCR-amplify the expression cassette and combine it with pAdenoX-PRLS in an In-Fusion HD Cloning reaction. In-Fusion HD Cloning is fast, simple, precise and efficient, making Adeno-X Adenoviral System 3 the most advanced commercially available adenoviral gene delivery tool.

Constructing recombinant adenovirus with In-Fusion Cloning technology.DNA sequences can be rapidly transferred as PCR products to any pAdenoX vector using the In-Fusion cloning method. In this example, your gene of interest is amplified with 15 bp extensions that are homologous to the ends of the linearized adenoviral vector. The PCR product is then purified and mixed with the linearized adenoviral vector of choice in the In-Fusion reaction. Following the reaction, a portion of the mixture is transformed intoE. coli(Stellar Competent Cells) and screened. Once a PCR-positive clone is identified, the recombinant pAdenoX vector is amplified, purified, and subsequently linearized with the restriction enzyme PacI, then transfected into Adeno-X 293 cells for viral rescue and amplification. Adeno-X GoStix can be used to determine the status of adenovirus rescue.

The universal Adeno-X adenoviral expression systems contain vectors that lack a promoter and polyA signal in the cloning site.You can either clone an expression cassette from a pre-existing construct into the vector (Panel A) or create a new one using multiple fragment In-Fusion HD Cloning (Panel B).