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. Pharmacodynamics of chloroquine Plaquenil and mucinex Chloroquine lysosomal membrane permeabilization May 07, 2001 Transfection efficiency was affected by the concentration of lysosomotropic agents. The marked transfection enhancement was observed when the amount of sucrose increased from 5 to 500 mM and the optimum transfection efficiency was found at 500 mM in all cell lines tested Fig. 2 a–c. Mar 11, 1983 Chloroquine treatment of rodent cells during the first hours of polyoma DNA transfection increase the fraction of cells expressing viral functions. The effect has been observed after DNA absorption using both the DEAE-dextran and calcium phosphate coprecipitation. PEI transfection protocol 24-well protocol 1. Preparation of the transfection solution For one well of a 24 well plate 100,000 cells, 2µg of DNA and 10µl of PEI 25KD, 10uM=250ug/ml. NB These quantities and volumes should be scaled up according to the number of wells to be transfected. • Dilute 2µg of DNA in 50µl of NaCl 150mM. Vortex. 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 Polyethylenimine - Wikipedia, High efficiency polyoma DNA transfection of chloroquine. Hydroxychloroquine tablet uses in tamilHydroxychloroquine side efectsCelebrex and plaquenilHydroxychloroquine will it help ringwormPurchase chloroquine tablets in uk 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. Chloroquine use in transfection. - Tissue and Cell Culture. PEI transfection protocol. Addgene General Transfection. 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. Add 22 l of 25mM chloroquine to each 15cm dish for 10cm plate 7.8 l of chloroquinei.e. final concentration = 25 M. The cells should be transfected within 5-10’ of adding the chloroquine. 7. For 15cm plates, add 1.69ml of the transfection buffer to the DNA-CaCl2 solution by “bubbling”. Chloroquine, which buffers lysosomes, enhanced the transfection efficiency of N-quaternized PEIs and polylysine by 2-3-fold. In contrast, chloroquine did not improve the transfection efficiency of PEI. The measured average pH environment of PEI vectors was 6.1, indicating that they successfully avoid trafficking to acidic lysosomes.