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. Chloroquine phosphate for lupus Chloroquine phosphate injection spc Plaquenil tendon pain Plaquenil 400 mg 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. HEK293F suspension culture transfection protocol Moremen lab, 12/15/11 Transient Transfection of HEK-293F Suspension Cultures using PEI Cells are grown in suspension on a platform shaker in a humidified 37°C CO The optimal amount of DNA to use in the transfection will vary widely depending upon the. PEI and dendrimers. medium with serum containing 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. Using pei vs chloroquine transfection Transfection of naked DNA - Jagiellonian University, Transfection of 293F Mammalian Cells using PEI Is chloroquine related to malaroneDoes hydroxychloroquine make you gain weightHydroxychloroquine methotrexate sulfasalazine azathioprine lefludomideChloroquine injection side effects As shown in Fig. 1b, the titers of lentiviral vectors produced using PEI-mediated transfection were only slightly lower than those involving the CaP-based transfection method. Also, the PEI-based method resulted in robust titers 10 7 TU/ml even though the transfection conditions had not been optimized at this point. It was also interesting. Simplified lentivirus vector production in protein-free media using.. Transfection Methods Overview - Bio-Rad. Transfection Guide Overview of Transfection Methods Promega. For CaPO3, I use chloroquine, which inhibits DNA degradation by lysosomes by raising their pH - but this is toxic to the cell, so media must be changed after 6-8 hr. PEI also prevents DNA. Gene transfer using a chemical carrier is called transfection. Transfection of DNA or RNA into animal cells has to be restricted to non-viral systems that transiently open pores into the cell membrane to permit the entry of nucleic acids. Phoenix - Helper dependent protocol. Preparing Retrovirus using Phoenix Lines. This is one of two transfection protocols you can find in this web page set. About 5 minutes prior to transfection, add chloroquine to each plate to 25uM chloroquine stock is 50 mM; for 3 mL media + 1 mL DNA, add 2u1. Chloroquine acts to inhibit lysosomal.