Welcome guest, Sign in



Thanks for your further question/comment. It has been sent to the author(s) of this protocol. You will receive a notification once your question/comment is addressed again by the author(s).
Meanwhile, it would be great if you could help us to spread the word about Bio-protocol.


[Bio101] Lentivirus Infection

Molecular Biology > DNA > Transfection
Author: Nabila Aboulaich
2/20/2011, 19035 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.37


Materials and Reagents

  1. Cells
  2. Polybrene (hexadimethrine bromide) (Sigma-Aldrich, catalog number: H9268 )
  3. Puromycin
  4. Human or mouse cell line and appropriate growth media reagents required for cell-based assay
  5. Polybrene appropriated antibiotics for selection purpose


  1. Tissue culture Incubator
  2. 6 cm tissue culture plates (Thermo Fisher Scientific)


Note: Lentiviral infections should be optimized for each cell line and cell-based assay. For example, the following parameters should be tested before starting large-scale infections to determine the optimal conditions for a given experiment:

  1. Cell seeding density.
  2. Amount of lentivirus.
  3. Puromycin concentration.
  4. Timecourse
  1. Seed cells at appropriate density in 6 ml in 6 cm plates.
    1. Adherent cells: seed 1 day prior to infection.
    2. Suspension cells: seed day of infection in media containing polybrene.
  2. Add virus to cells:
    1. (Adherent cells): Remove growth media and add fresh media containing polybrene. Alternatively, remove a portion of the growth media and supplement with media containing polybrene. Adjust volumes and polybrene concentration to achieve the correct final polybrene concentration (8 μg/ml).
  3. Viral infection:
    1. Incubate cells overnight.
    2. Change media 24 h post-infection. Remove media and replace with 6 ml fresh growth media. If antibiotics selection is desired, use fresh growth media containing antibiotics.
      Note: Puromycin concentration should be optimized for each cell line; typical concentrations range from 2-5 μg/ml.
  4. Incubate cells, replacing growth media (with antibiotics, if desired) as needed every few days. Incubation periods are highly dependent on the post-infection assay.
    Note: All lentiviral procedures should be carried out in accordance with biosafety requirements of the host institution.

How to cite: Aboulaich, N. (2011). Lentivirus Infection. Bio-protocol Bio101: e37. DOI: 10.21769/BioProtoc.37; Full Text

Share Your Feedback:

  • Add Photo
  • Add Video

Bio-protocol's major goal is to make reproducing an experiment an easier task. If you have used this protocol, it would be great if you could share your experience by leaving some comments, uploading images or even sharing some videos. Please login to post your feedback.

Ask the Authors:

  • Add Photo
  • Add Video

Please login to post your questions/comments. Your questions will be directed to the authors of the protocol. The authors will be requested to answer your questions at their earliest convenience. Once your questions are answered, you will be informed using the email address that you register with bio-protocol.
You are highly recommended to post your data (images or even videos) for the troubleshooting. For uploading videos, you may need a Google account because Bio-protocol uses YouTube to host videos.

Login | Register
How to cite
Twitter Twitter
LinkedIn LinkedIn
Google+ Google+
Facebook Facebook