This Retro-X qRT-PCR Titration Kit and a real-time PCR instrument can be used to rapidly quantify the virion content of MMLV-based retroviral supernatants. The viral genome copy number in a supernatant sample is determined by first purifying the viral RNA, and then comparing the RT-PCR threshold cycle (C_t) of an appropriate dilution of purified viral RNA to sample curve C_t values that are derived from a calibrated, serially diluted control viral RNA template.

This kit provides a complete retroviral vector packaging system for use with any retroviral vector. It is designed to produce virus that can infect dividing cells from a broad range of mammalian and nonmammalian cell types. The GP2-293 Packaging Cell Line contains only the MoMuLV gag and pol genes. The viral envelope portion of the packaging function (env gene) is supplied by transiently co-transfecting one of the Retro-X Universal Packaging Vectors (p10A1, pAmpho, pEco, or pVSV-G) with the retroviral vector. The pQCLIN Retroviral Vector (included in the vector set) can be used as a positive control to constitutively express β-galactosidase in transduced mammalian cells.

The Retro-X Concentrator is a complete reagent for the concentration of infectious retroviral vector particles. This reagent provides a scalable alternative to ultracentrifugation for viral particle concentration. Vector supernatants can be concentrated 10–100 fold, depending upon the volumes and envelope used.

RetroNectin reagent is a 63-kD fragment of recombinant human fibronectin fragment (also referred to as rFN-CH-296) that enhances the efficiency of lentiviral- and retroviral-mediated gene transduction. This is particularly important for hematopoietic cells and other hard-to-transfect cell types. When coated on the surface of containers such as culture dishes, petri dishes, flasks or bags, RetroNectin significantly enhances virus-mediated gene transduction into mammalian cells. This enhanced transduction is thought to result from the colocalization of virus particles and target cells on the molecules of RetroNectin. Viral particles bind RetroNectin via interaction with heparin-binding domain II, and target cells bind mainly through the interaction of the cell surface integrin receptor VLA-4 with the fibronectin CS1 site.

This product is a plasmid vector for producing a high expression efficiency retrovirus vector. By transfecting this vector into an appropriate packaging cell line, the vector transiently or stably expresses transcribed products containing the virus-packaging signal (Ψ), target gene and selective marker. This vector possesses LTR and Ψ (viral packaging signal), but not the structural genes necessary for particle formation and replication (sequences gag, pol, and env). Because this vector contains an intron with high splicing capacity, it yields high transcription efficiency. pMEI-5 Neo DNA (Cat.# 3655) contains the neomycin resistance gene as a selective marker. Gene expression two to eight-fold higher than pDON-Al-2 series can be expected.