Chloroquine dna transfection

Discussion in 'Generic Chloroquine Online' started by ValidData, 12-Mar-2020.

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    Chloroquine dna transfection


    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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    Typically, DNA is suspended in sterile water or TE buffer to a final concentration of 0.2–1 mg/ml The optimal amount of DNA to use in the transfection will vary widely depending upon the type of DNA, transfection reagent/method, target cell line, and number of cells Genetic Material DNA Quality and Quantity Factors Affecting We describe a highly efficient stable gene transfection procedure for Chinese hamster ovary CHO cells using a modification of the calcium phosphate-DNA coprecipitation method. We have found that treatment of CHO cells with chloroquine increases the efficiency of gene transfer by up to 20-fold from approx. 0.01% to approx. 0.2% when transfection is done using the pSV2-neo plasmid. The. Chloroquine is a member of quinolone family and is a weak intercalating agent. Chloroquine is used for treating amebiasis, rheumatoid arthritis, discoid and systemic lupus erythematosus. Application DNA intercalator. Also used to increase transfection efficiency. Chloroquine diphosphate salt has been used • in in vitro antiplasmodial assays

    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 dna transfection

    Helper Dependent Protocol - Stanford University, High-efficiency stable gene transfection using chloroquine.

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  7. Containing 100 µM chloroquine with-out serum were added to each well, followed by incubation for 90 min in 5% CO2 at 37°C. Cells were washed once in PBS and cultured in R10 for a total of 24 h from the start of transfec-tion. LIPOFECTIN Transfection For each transfection with LIPO-FECTIN, 1 × 106 cells were distributed in

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    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. After cells chloroquine-treated or untreated have been exposed to the calcium phosphate–DNA coprecipitate in growth medium for 2–6 h, remove the medium by aspiration and wash the. 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. Add 10µg DNA to ddH 2 O 1095 µl total in 15-ml sterile tube, then add 155 µl 2M CaCl 2. When you are

     
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