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Home › Products › Gene function › Viral transduction › Lentivirus › Titration kits › Integrated copy number

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Integrated lentivirus copy number quantification

Determine the copy number of integrated lentiviruses (proviruses) present in transduced cells.

The Lenti-X Provirus Quantitation Kit allows you to rapidly determine the copy number of integrated lentiviruses (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 the calibrated control template or purified cellular genomic DNA.

The Lenti-X Provirus Quantitation Kit allows you to rapidly determine the copy number of integrated lentiviruses (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 the calibrated control template or purified cellular genomic DNA.

Determine precise effective lentivirus titers

By quantifying the number of lentiviruses that have integrated into the nuclear DNA of your target cells, you can precisely determine the real titer (i.e., effective titer) of your lentiviral supernatant stock and use this information to infect cells with greater precision and a more highly defined MOI (multiplicity of infection), allowing you to predict how many viral genomes will integrate into your cells. You can also use this method to calibrate other titration methods which measure virus content in packaging-cell supernatants but do not measure functional integrative titers in your target cells.

Predictable and consistent lentiviral transgene expression

When using lentiviral vectors to deliver transgenes into target cells, expression levels increase with increasing MOI because more lentivirus copies integrate into the nuclear DNA of the cells. Also, for a given virus prep, its effective MOI across different target cell lines can vary widely depending on each cell line’s individual susceptibility to lentivirus infection. The Lenti-X Provirus Quantitation Kit allows you to assess the results of your transductions in any cell line, independent of expression level, and can be used to ensure consistent transductions across different cell lines or when using different virus preparations.

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Cat. # Product Size Price License Quantity Details
631239 Lenti-X™ Provirus Quantitation Kit 200 Rxns USD $602.00

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.

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Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Measuring lentiviral copy number in target cells

Measuring lentiviral copy number in target cells
Measuring lentiviral copy number in target cells. Provirus content is determined using a rapid qPCR method.

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Lentiviral provirus copy number affects transgene expression levels in transduced cells

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.

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Equivalent virus amounts in different cell types result in different provirus (integrated lentivirus) copy numbers

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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631239: Lenti-X Provirus Quantitation Kit

631239: Lenti-X Provirus Quantitation Kit

Lenti-X GoStix Plus how-to videos Lenti-X GoStix Plus video protocols
Selection guides Lentiviral vector options
Learn about lentiviral transduction experiments Lentivirus learning center

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*

*The primers included in this kit are designed to amplify a conserved region of HIV-1 genome adjacent to the packaging signal. These primers are compatible with all lentiviral vectors sold by Takara Bio and most commonly used lentiviral vectors. 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.


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Lenti-X Concentrator

Lenti-X Concentrator video

Lenti-X Concentrator is a simple and fast method for concentrating any lentiviral stock—just mix your lentiviral supernatant with the concentration reagent, incubate, and spin the mixture in a standard centrifuge. Watch the video to see how it works.

Watch Concentrator video Lentiviral packaging overview

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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