PMC:6218659 / 80217-81175 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/6218659","sourcedb":"PMC","sourceid":"6218659","source_url":"https://www.ncbi.nlm.nih.gov/pmc/6218659","text":"Transfection Protocol\nTransfection reagents and medium were prepared fresh every time. A PEI (Polyethylimine) stock solution was first prepared (25 kDa, linear -100 μg/mL). PEI was added in MilliQ water and HCl (aq) was then added drop-wise with just enough to help solubilize the PEI. The PEI solution was filtered through 0.2 um membrane in laminar sterile hood. PEI solution was set aside. The DNA stock solution was added to serum-free DMEM medium at room temperature to make final DNA-DMEM concentration of 4 μg/mL. PEI solution was then added to DMEM-DNA solution to make a final concentration of PEI of 4.6 μg/mL. The PEI-DNA transfection medium was gently stirred at room temperature for 20 minutes before use. For the transfection step, the DNA-PEI transfection was added first and placed in the incubator (37 °C with 5% CO2) for 10 minutes and then DMEM with 2% FBS was added. The DNA-PEI medium and DMEM with 2% FBS medium were added in 1:3 ratio.","divisions":[{"label":"Title","span":{"begin":0,"end":21}}],"tracks":[]}