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 (Ct) of an appropriate dilution of purified viral RNA to sample curve Ct values that are derived from a calibrated, serially diluted control viral RNA template.
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.
