Lenti-X Provirus Quantitation Kit
Lenti-X Provirus Quantitation Kit
Brand: Takara Bio.
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Lenti-X Provirus Quantitation Kit
The Lenti-X Provirus Quantitation Kit allows you to rapidly determine the provirus content of cells that have been transduced with lentivirus. The kit provides components to perform real-time quantitative PCR of integrated proviruses in genomic DNA, and can be used to determine the copy number of proviruses present in a mixed or clonal population of transduced cells. The kit includes a genomic DNA purification kit, and all the reagents required to conduct 200 TB Green-based qPCR reactions using either a calibrated control template or purified cellular genomic DNA.
Overview
- Predict consistent transgene expression levels between experiments
- Compare transduction data across multiple cell lines
- Ensure consistent copy number between experiments or across different cell lines
- Reagents for 10 genomic DNA purifications and 200 TB Green qPCR reactions
Measuring lentiviral copy number in target cells
Measuring lentiviral copy number in target cells. Provirus content is determined using a rapid qPCR method.
Lentiviral provirus copy number affects transgene expression levels in transduced cells
Lentiviral provirus copy number affects transgene expression levels in transduced cells. HT1080 cells were infected at either high or low MOI using an AcGFP1 lentiviral expression vector. 72 hr after infection, the cells were subjected to puromycin drug selection. One week later, stably transduced clones were selected for expansion and analysis. AcGFP1 expression levels were determined using flow cytometry, and provirus content in genomic DNA was determined using the Lenti-X Provirus Quantitation Kit. The figure represents data pooled from two groups having either low copy number (>3; n = 8) or high copy number (>9; n = 7). Higher provirus contents were associated with higher expression levels.
Equivalent virus amounts in different cell types result in different provirus (integrated lentivirus) copy numbers
Equivalent virus amounts in different cell types results in different provirus (integrated lentivirus) copy numbers. Equivalent amounts of a single lentiviral stock (100 µl) were used to infect either HT1080, HeLa, and HEK 293 cells. 72 hr after infection, the cells were subjected to drug selection with puromycin. One week later, the provirus copy numbers of the stably transduced cell populations were determined using the Lenti-X Provirus Quantitation Kit.
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Measuring lentiviral copy number in target cells
Measuring lentiviral copy number in target cells. Provirus content is determined using a rapid qPCR method.
Lentiviral provirus copy number affects transgene expression levels in transduced cells
Lentiviral provirus copy number affects transgene expression levels in transduced cells. HT1080 cells were infected at either high or low MOI using an AcGFP1 lentiviral expression vector. 72 hr after infection, the cells were subjected to puromycin drug selection. One week later, stably transduced clones were selected for expansion and analysis. AcGFP1 expression levels were determined using flow cytometry, and provirus content in genomic DNA was determined using the Lenti-X Provirus Quantitation Kit. The figure represents data pooled from two groups having either low copy number (>3; n = 8) or high copy number (>9; n = 7). Higher provirus contents were associated with higher expression levels.
Equivalent virus amounts in different cell types result in different provirus (integrated lentivirus) copy numbers
Equivalent virus amounts in different cell types results in different provirus (integrated lentivirus) copy numbers. Equivalent amounts of a single lentiviral stock (100 µl) were used to infect either HT1080, HeLa, and HEK 293 cells. 72 hr after infection, the cells were subjected to drug selection with puromycin. One week later, the provirus copy numbers of the stably transduced cell populations were determined using the Lenti-X Provirus Quantitation Kit.
