- Home
- Protocols
-
Lentivirus Production
Published: Mar 5, 2011 DOI: 10.21769/BioProtoc.39 Views: 51508
PDF 
Ask a question
Abstract
Lentivirus is a common tool used to introduce a gene into mammalian or other animal cells.This protocol is to produce lentivirus stocks from hairpin-pLKO.1 plasmid.
Materials and Reagents
- 293T packaging cells (ATCC)
- Hairpin-pLKO.1 vector (GE Healthcare Dharmacon)
- Packaging vectors: 2nd generation packaging plasmids containing gag, pol, and rev genes (pCMV-dR8.91 or pCMV-dR8.74psPAX2) and envelop plasmid (pMD2.G) (Addgene)
- Lipofectamine 2000 (Life Technologies, Invitrogen™)
- OptiMEM serum free media (Life Technologies, Invitrogen™)
- DMEM (Life Technologies, Invitrogen™)
- Fetal Bovine Serum (FBS) (Life Technologies, Invitrogen™)
- Pen/Strep (Life Technologies, Invitrogen™)
- 6 cm cell culture plates (Thermo Fisher Scientific)
- 45 μm filter (Thermo Fisher Scientific)
- Storage tubes
- Seeding media (see Recipes)
- Harvest media (see Recipes)
Equipment
- Tissue culture incubator (37 °C, 5% CO2)
- Centrifuges
Procedure
Category
Microbiology > Microbial biochemistry > RNA
Cell Biology > Cell isolation and culture > Cell growth
Molecular Biology > RNA > Transfection
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this article to respond.
Tips for asking effective questions
+ Description
Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.
Post a Question
