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. Does plaquenil have to build up in your system Hydroxychloroquine multiple sclerosis Chloroquine: mechanism of drug action and resistance in plasmodium falciparum 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. 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. Add transfection reaction to this media, and incubate o/n JoVE. Pro-Tip* The ratio of ug DNAug PEI needs to be empirically determined. Once a batch of PEI is prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the cells 1-2 days after transfection to determine what ratio gives the highest percentage of GFP positive cells. 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 pei transfection Chloroquine to improve transfection - Cell Biology, Glycerol shock or chloroquine? - Transfection and Transduction Chloroquine resistant malaria pptHydroxychloroquine versus sulfasalazinePlaquenil mefloquine Chloroquine diphosphate 52 mg Deionized, distilled H 2 O 1 mL Dissolve 52 mg of chloroquine diphosphate in 1 mL of deionized, distilled H 2 O. Sterilize the solution by passing it through a 0.22-µm filter. Store the filtrate in foil-wrapped tubes at −20°C. Chloroquine Diphosphate 100 mm for Transfection. Addgene General Transfection. Transfection Guide Overview of Transfection Methods Promega. Antibacterial agents were used in growth medium during transfection Do not use antibiotics such as chloroquine, penicillin, or streptomycin in growth medium because during transfection, cells are more permeable to antibiotics, which may cause toxicity. Transfection reagent stored improperly We recommend storing transfection reagents at 4°C. The efficient miRNA delivery as higher content of chloroquine caused increased endosomal escape 22. In another study con-ducted by Yang Hu et al. in the presence of 75 μM concentration of CQ, polyethyleneimine PEI/pDNA polyplex at its optimal N/P showed 2.2 times higher transfection efficiency than PEI-pDNA Optimized PEI-based transfection reagent for scalable virus production. With its unique property to efficiently condense several plasmids of different sizes, PEIpro ® transfection reagent is ideal for optimal co-transfection of several plasmids containing the gene of interest and necessary viral components to produce full recombinant virions.