Retrovirus titration by qRT-PCR
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.
Overview
- Measure retrovirus titers using TB Green-based, real-time PCR
- Retrovirus copy numbers are determined by comparing the sample's Ct to a standard curve derived from a calibrated control template
- Confirm the success of your retroviral packaging reaction to avoid wasting time with your experiments
- Perform consistent experiments, control copy number, and hence expression levels
- Ensure sufficient MOI so that 100% of your cells are transduced
- 200 qRT-PCR reactions* are included, and 10 different virus preps can be purified
*The primers included in this kit are designed to amplify a conserved region of the MMLV genome adjacent to the packaging signal. These primers are compatible with all MMLV vectors sold by Takara Bio (e.g., Q-series and L-series vectors) and most commonly used retroviral vectors. However, vectors carrying a significantly modified packaging signal, like the one carried by MSCV vectors, would be incompatible with the kit. If you are unsure if this kit is compatible with your vector, please send your vector sequence to Technical_Support@takarabio.com, and our support team will be happy to confirm primer compatibility.
More Information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
Retroviral product selection guide
Choose from a wide variety of retroviral vectors, as well as retroviral packaging systems and cell lines, products for titrating and concentrating retrovirus, and retroviral transduction enhancers.
Selection guide Recombinant virus comparisonTakara Bio USA, Inc.
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