DNA quantification and normalization

∗ Incubate the plates for 2 to 5 minutes at room temperature, protected from light,before the analysis on the 7900 HT Sequence Detection System.Analyze the PicoGreen plates in the 7900 HT Sequence Detection System∗ Change to the block module intended for 96 well reaction plates.∗ Make a new SDS document.Assay: allelic discriminationContainer: 96 wells clear plateMarker: PicoGreen, with the detectors SYBR and VIC. Passive reference: -none-∗ Connect to the instrument and press “Post Read”. Analyze your plate and export theresult table.Calculate the concentrations of the samples in excel∗ Open the result table exported from SDS in excel, and save it as an excel-file.∗ The PicoGreen fluorescence is most often presented in the “Allele Y Rn” column,double check this with the SDS result plot. Make a dot diagram including the knownCeph concentrations (Y-axis: PicoGreen fluorescence, X-axis: known DNAconcentration). Insert trend line and show equation.∗ Calculate with the help of the equation the sample concentration. Remember to dividethe concentration with 1,5 since this was the volume DNA transferred to thePicoGreen plate.Make an import file for normalization on Biomek FX∗ Use the template “Mall DNAConc_import file normalization” (Analysdata server >PicoGreen folder). Save it as your own and store it in your folder in Externa konsulter.Biomek FX and 7900 HT090116/CN