Lentivirus integration site analysis kit
Identify the exact site of integration of recombinant lentiviral vectors using the Lenti-X Integration Site Analysis Kit. The replication life cycle of a lentivirus results in integration of a DNA copy of the lentiviral RNA genome into the host genome. The chosen integration site has important consequences for both the expression of your transgene and the phenotype of the host cell.
Identify the exact site of integration of recombinant lentiviral vectors using the Lenti-X Integration Site Analysis Kit. The replication life cycle of a lentivirus results in integration of a DNA copy of the lentiviral RNA genome into the host genome. The chosen integration site has important consequences for both the expression of your transgene and the phenotype of the host cell. The lentiviral integration site analysis kit uses GenomeWalker technology to identify those sites so that they can be sequenced and analyzed.
The GenomeWalker procedure
- Genomic DNA from transduced clones or populations is digested with blunt-end cutting restriction enzymes.
- The GenomeWalker adaptor is ligated to the blunt ends of the genomic DNA library.
- A primary PCR amplifies those fragments from the library that contain both an adaptor and a lentiviral sequence.
- A secondary ‘nested PCR’ generally produces a single, major PCR product that can be sequenced to determine the integration site. Multiple bands can result from multiple integrations sites, and each can be purified and sequenced separately.
What is in the Kit?
The kit provides all the components and controls needed to isolate and PCR amplify the lentiviral integration sites for three cell clones, including:
- NucleoSpin Tissue kit to purify high-quality genomic DNA from transduced mammalian cells
- Three blunt-cutter restriction enzymes, ligase, GenomeWalker adaptor, and PCR primers
- Advantage 2 PCR kit for long-distance amplification from genomic DNA
- NucleoSpin Gel and PCR Clean-Up kit for isolating the amplified band(s)
Overview
- Identify sites of lentiviral integration into the host genome
- Simple determination of lentivirus integration sites using GenomeWalker technology
- Compatible with most commercially available lentivirus vectors
- Complete system including all the necessary enzymes, primers, controls, PCR kit, genomic DNA purification kit, and gel purification kit
- Retroviral version also available for detecting integration sites for MMLV and MSCV retroviruses
- Identify sites of lentiviral integration into the host genome
More Information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the kit's Components List.
Lentiviral product selection guide
Takara Bio offers a broad range of research tools for gene delivery using lentiviral vectors, including a selection of vector systems, packaging mixes, and titration kits. Use our selection guide for a brief description of the products and links to further information.
Lentiviral products guide Learning centerTakara Bio USA, Inc.
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