Preparing 48-Well Plate FeedersIn preparing 48-well plates, a dilution must be made from the regulardensity of 3.5 x 105 cells/ml. The required density for 48-well plates is: 8.2 x 104 cells/ml (650 ml/well or 31.2 mls/plate).Example:For a total of 5 x 48-well plates, the calculation is:5 plates x 31.2 mls = 156 mls + 10 mls Extra = 166 mls (Total)166 mls x 8.2 x 104 cells/ml = 1.36 x 107 cells (Total)1.36 x 107 cells : 3.5 x 105 cells/ml = 38.86 mlsDilution: 38.9 mls (of cell sUSPension@3.5x105cells/ml)+127.1mlsMedia.Takeacountatthispoint,toverifythecelldensityof8.2x104cells/ml.Plate-outthisfeedermedia(dilution)intothe48-wellplates.REMEMBERTOUSEPLATESTHATHAVEBEENGELLED,2HOURSPRIORTHIS.Dispense650mloffeedermedia/well(31.2mls/plate).Mixplatestodistributecellsuniformlyandincubate.No.ofPlatesTotal#ofCellsDilution(mls)TotalVol.(mls)5plates1.36x107cells38.9+127.1166mls10plates2.64x107cells75.0+247.032215plates3.92x107cells112.0+366.047820plates5.20x107cells148.5+485.5634Feeders/STOMedia:DMEM,7%FCS,1XGPS.FromtheLaboratoryofDr.AllanBradleyBaylorCollegeofMedicine,Houston,Texas
