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Home › Learning centers › Gene function › Viral transduction › Lentivirus › Lentiviral workflow

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Selection guides Virus kits: product finder tool
Tools for recombinant lentivirus production and evaluation Lentivirus product catalog

Lentiviral workflow

Lentiviral gene delivery tools

Use the typical lentivirus production workflow depicted below to find products, learning resources, and technical information related to each step of your experiment. For an overview of all available viral transduction systems, visit our viral transduction learning center.

Choose your vector

When choosing a plasmid backbone, you have many features to consider for expression of your gene of interest. Constitutive or inducible expression? Untagged or fluorescently labeled? Explore available options to find the best vector for your experimental aim.

Clone your gene

In-Fusion cloning is ideal for inserting your fragment into a lentiviral vector—just one quick reaction allows you to recover your final construct.

Learn how In-Fusion Cloning was used to quickly clone PCR fragments into a lentiviral vector that had only one suitable restriction site for subcloning.

Package your virus

The most critical factor for successful lentiviral transduction is viral titer. Start with our fourth generation packaging system and obtain between 107and 108 infectious units per ml, 25 times what other popular systems generate.

Titrate your virus

It is important to have an accurate estimation of infectious forming units (IFU) in your packaging supernatant. This measurement will determine how much supernatant is used to achieve a desired MOI—and ensure successful gene delivery. There are 4 titration kit options available from Takara Bio:

  1. Need a fast answer? Lenti-X GoStix take only 10 minutes to assess lentivirus titer and determine whether your supernatants are ready for harvesting.
  2. Use an accurate ELISA-based protocol to measure p24 capsid protein content in your supernatant.
  3. Use qRT-PCR to determine the viral genome content in your viral supernant from a calibrated RNA standard curve.
  4. Quantify the number of lentiviruses that have integrated into the nuclear DNA of your target cells (provirus) using a green intercalating dye-based qPCR reaction.

For more information see the lentiviral titration kit selection guide.

Concentrate your virus

If your IFU/ml titer is low, Lenti-X Concentrator can be used to concentrate your virus up to 100-fold and improve gene delivery. This concentration method is scalable, easier, and faster than ultracentrifugation.

Transduce your target cells

Transduction problems can be frustrating and waste valuable resources—inhibitors in your media or difficult cell types can hinder lentiviral gene delivery. Learn how to navigate transduction troubles.

Want to avoid Polybrene and speed up transduction? Bring virus particles from your lentiviral packaging supernatant and cultured cells together with charged magnetic beads. Some cell types, such as hematopoietic cells, show very low transduction efficiency even with high-titer lentivirus. RetroNectin reagent promotes co-localization of virus and cells to dramatically enhance transduction efficiency.

Analyze integration

The life cycle of lentivirus results in integration of a copy of the lentiviral 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 specific integration site.


Related product pages

  • Viral product finder tool
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Capturem Trypsin provides rapid, efficient, and complete digestion of protein samples, allowing an uninterrupted mass spectometry workflow at room temperature for downstream protein analysis. This product utilizes our novel Capturem technology in a spin column format with membrane-immobilized trypsin. Capturem Trypsin Columns may be used to completely digest protein samples in less than a minute with digestion efficiencies (protein coverage) comparable to or better than those obtained using in-solution trypsin digestion.

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