Vector Protocols

AAV Vector Production and Purification Protocol Protocol is used to propagate and purify AAV vectors for experiments both in vitro and in vivo. Includes: Principles of the Triple Plasmid Transfection System; Plasmids; Transfection and Extraction of Virus; Purification of the AAV vector.

Adherent Cell Types Infection Protocol Protocol for infection for adherent cell types.

Blunt-ended Cloning into Plasmid Vectors Protocol Protocol for blunt-ended cloning into plasmid vectors.

Cloning siRNA Duplexes into pSUPER or pSUPER Retro Vectors Protocol Protocol for cloning siRNA duplexes into pSUPER or pSUPER retro vectors.

Directional Cloning into Plasmid Vectors Protocol Directional cloning requires that the plasmid vector be cleaved with two restriction enzymes that generate incompatible termini and that the fragment of DNA to be cloned carries termini that are compatible with those of the doubly cleaved vector.

In Vivo Cloning with Mu Protocol Protocol for in vivo cloning with mu.

Lineage Analysis Using Retroviral Vectors Protocol Protocol describes the use of replication-incompetent retroviral vectors for the analysis of lineal relationships in developing vertebrate tissues.

Phosphatase Treatment of Linearized Vector Protocol To minimize self-ligated vector in your transformation, treat your linearized vector with a phosphatase to remove the 5' phosphates necessary for ligation. This should improve the percentage of colonies with inserts.

Transfection/Retroviral Infection pCL-Eco+Gene of Interest into 293T Cells Infecting HC11 Cells Protocol for the transfection/Retroviral Infection pCL-Eco+Gene of interest into 293T cells infecting HC11 cells. Includes: Split 293T cells for transfection; Transfect 293T cells; Split HC11 cells for infection; Infect target (HC11) cells.

Viral Vector Handling Information Information on how to handle viral vectors. Includes: Adenovirus;  MoMLV retrovirus; MoMLV-based Retroviral Vectors: Base classification BSL-1.