Chloroquine lentivirus production

Discussion in 'Canadian Ed Drugstore' started by Ivanilho, 07-Mar-2020.

  1. SmoRodina XenForo Moderator

    Chloroquine lentivirus production

    Transfections allow for transient expression of a gene of interest in a target cell line and can be useful for short term studies of protein function. We specifically use this protocol with Lenti-X 293T cells, a cell line optimized for production of lentiviral vectors.

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    MM chloroquine diphosphate Dissolve 0.129 g of chloroquine diphosphate salt in 10 mL of sterile water. Filter sterilize through a 0.22 μm filter. Aliquot 50-100 μL and store at -20 ℃. Aliquots can be thawed and stored at 4 ℃ prior to use. Thawed aliquots should be discarded after 1-2 months. Chloroquine is an inhibitor of the lysosomal degradation of the DNA which is taken up by the cells, so as leelee said, transfection should have been successful, albeit at a slightly lower level than if you would have added the chloroquine. In my normal transfections, I never add chloroquine and get high efficiencies nevertheless. Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors Robert H Kutner1, Xian-Yang Zhang1 & Jakob Reiser1,2 1Gene Therapy Program, Department of Medicine, Louisiana State University Health Sciences Center, 533 Bolivar Street, New Orleans, Louisiana 70112, USA. 2US Food

    Last Upload: June 10, 2016 Day 0: Seed Lenti-X 293T cells (this cell line is optimized for production of lentiviral vectors) Day 1 (pm): Transfect Cells Day 2 (am): 18h post transfection - Remove media, replace with fresh media Day 3 or more (am): Observe fluorescence, harvest cells, or perform your experiment *Pro-Tips* Different brands and lots of FBS can promote or inhibit transfection. This approach can be adapted for different cell lines and different transfection reagents.

    Chloroquine lentivirus production

    Production, concentration and titration of pseudotyped HIV., Chloroquine use in transfection. - Tissue and Cell Culture

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  7. Which concentrations of Chloroquine and Na-Butyrate are suitable for Lentivirus production in HEK293T cells. Chloroquine concentrations range from 25nM to 2mM final concentration in the plate.

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    Lentivirus Production. Lentiviral preparations are produced in a 293T cell line using the 2nd generation psPAX2 and pMD2. G packaging system developed by the Trono culture medium OptiPro SFM containing lentivirus is first cleared by low speed centrifugation and then by filtration through a 0.45 µm polyethersulfone membrane. The provided production medium does not contain chemicals used to add lentivirus production such as sodium butyrate, caffeine, or chloroquine. So, lentivirus in this medium can be directly used for most of applications. 3. Reagents for purification and long-term storage are included. 4. Chloroquine is an anti-malaria medicine that works by interfering with the growth of parasites in the red blood cells of the human body. Parasites that cause malaria typically enter the body through the bite of a mosquito.

  8. renatko1991 Guest

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