This Lenti-X qRT-PCR Titration Kit and a real-time PCR instrument can be used to rapidly quantify the virion content of HIV-1-based lentiviral 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
- Confirm the success of your lentiviral packaging reaction
- Perform consistent experiments
- Ensure sufficient MOIs so that 100% of your cells are transduced
- Control copy number and hence expression levels
- 200 qRT-PCR reactions are included*
- Purify 10 different virus preps
