The Retro-X qRT-PCR Titration Kit provides a fast and simple method for measuring retrovirus titer. The kits use a quick RNA purification step before quantifying the number of retroviral genome copies using qRT-PCR and TB Green technologies.
The Retro-X qRT-PCR Titration Kit provides a fast and simple method for measuring retrovirus titer. The kits use a quick RNA purification step before quantifying the number of retroviral genome copies using qRT-PCR and TB Green technologies.
Rapid determination of retroviral titer
Whereas traditional titration methods require up to 10 days to complete, these titration kits require only 4 hours and work with all MMLV-based retroviral vectors (not MSCV). Using qRT-PCR dramatically shortens the time interval between viral harvest and target-cell infection, allowing you to perform both on the same day.
Achieve consistent retroviral transductions
A major advantage of retroviral gene-delivery systems is the ability to control the level of gene expression by adjusting the multiplicity of infection (MOI) to dictate the number of viral genomes that enter your target cells. These titration kits allow you to make a rapid correlation between the retroviral copy number and retroviral infectivity in your cell type and ultimately allow you to control the expression level between experiments.
How it works
The qRT-PCR procedure entails subjecting serial dilutions of your sample and a control RNA of known copy number (provided) to one-step qRT-PCR. The retroviral copy number contained in your sample can then be determined by comparing its Ct value to the standard curve.
RNA purification and DNase I treatment steps are critical to the protocol so even viruses packaged using transient transfections, which always contain residual plasmid DNA, can be accurately quantified using this kit.