This procedure can be modified for alternative packaging cell lines or transfection reagents. Once produced, lentivirus can be used for a variety of downstream applications such as stable-cell line generation. Plaquenil 200 mg sanofi aventis Teva-chloroquine phosphate Plaquenil and cns Plate cells the night before to give 60-80% confluence at the day of transfection. 2. One hour prior to the transfection, change to medium containing 25µM chloroquine from x1000 stock in PBS, stored at -20°C. Volume should be 10 ml per dish. Chloroquine can be omitted, but increases efficiency about x2 3. Test a variety of brands and lots of FBS to find one suitable with your protocols. FBS can be purchased already heat inactivated or it can be inactivated in the lab by heating to 56 ℃ for 30 minutes. 25 mM chloroquine diphosphate Dissolve 0.129 g of chloroquine diphosphate salt in 10 mL of sterile water. The following are the protocols I've gathered *4-6 hours after transfection reaction has been applied to cells, gently remove media and replace with 10% glycerol or DMSO for 2-3 minutes. Remove and feed with complete media various references. *1 hour before transfection, replace cDMEM with cDMEM + 25 mM chloroquine. Test a variety of brands and lots of FBS to find one suitable with your protocols. Last Upload: August 26, 2016 *Pro-Tip* Different brands and lots of FBS can promote or inhibit transfection. Chloroquine transfection protocol Improve Lentiviral Production Using Lipofectamine 3000., Addgene Lentivirus Production Protocol Do prednisone and plaquenil work on ankylosing spondylitisPlaquenil muscle weaknessGive the synthesis of chloroquine May 07, 2001 Transfection efficiency in fibroblasts showed a linear increase 3–6-fold as the addition of chloroquine increased from 10 to 100 μM compared to plasmid alone Fig. 2 a–c. The maximum gene expression twofold increase compared to lipofectamine alone was observed in CHO cells when the cells were transfected with lipofectamine in the presence of chloroquine Fig. 2 b. Enhanced plasmid DNA transfection with lysosomotropic agents.. Glycerol shock or chloroquine? - Transfection and Transduction. Protocol for Lentivial Vector LV Production 2nd Generation.. The initial transfection is further improved by use of chloroquine. Stocks of ecotropic virus, collected as culture supernatant 48 hrs. post-transfection, can be stored at -80°C and used for infection of cell-lines in view of transformation and in vitro studies, or primary cells such as mouse bone marrow cells, that can then be used for transplant in our mouse model. I have different protocols for Lentivirus production using Lipofectamine transfection of HEK293T cells and now am confused about the concentration of Chloroquine and Na-Butyrate. Transfection Methods Reagent-Based Methods DEAE-Dextran Method Overview Solution A DNA ~1–5 µg/ml diluted into 2 ml of growth medium with serum containing chloroquine Solution B DEAE-dextran solution ~50–500 µg/ml Solution C ~5 ml of DMSO Solution D Complete growth medium 1 Add solution A to solution B, then mix gently.