1 2 3 4 Protocol Prepare Working Solution. Dilute the QuantiFluor® 1:200 in 1X TE buffer to make the QuantiFluor® example, add 10μl of QuantiFluor® Prepare Blank. Add 100μl of QuantiFluor® Note: If the Quantus™ Fluorometer was previously calibrated, you may not need to calibrate it again; do not prepare blank and standard samples, and skip Steps 2, 3 and 7. dsDNA Dye dsDNA Dye working solution. For dsDNA Dye to 1,990μl of 1X TE buffer, and mix. dsDNA Dye working solution and 100μl of 1X TE buffer to an empty 0.5ml PCR tube, and mix. This will be the blank sample used in Step 7. Protect tube from light. Prepare Standard. Dilute the DNA Standard to 2ng/μl by adding 2μl of the provided DNA Standard to 98μl of 1X TE buffer, and mix. Add 100μl of QuantiFluor® dsDNA Dye working solution, and mix. This will be the standard sample used in Step 7. Protect tube from light. Prepare Unknown(s). Add 100μl of unknown sample and 100μl of QuantiFluor® dsDNA Dye working solution to a 0.5ml PCR tube, and mix. Note: If the volume of the unknown DNA sample is less than 100μl, add 1X TE buffer to bring to a final volume of 100μl. For example, dilute a 1μl sample with 99μl of 1X TE buffer, then add 100μl of QuantiFluor® dsDNA Dye working solution for a total volume of 200μl for quantitation. 5 Incubate the prepared samples at room temperature for 5 minutes, protected from light. 6 Select the dsDNA protocol on the Quantus™ Fluorometer. 7 To Calibrate the Quantus™ Fluorometer, read the blank and standard samples in the Calibration screen, then select “Save”. 8 Enter the Volume of the unknown sample and desired concentration units. Note: This volume is the amount of sample that is added for the quantitation. For example, if 1μl of sample was mixed with 99μl of 1X TE buffer and then added to 100μl of QuantiFluor® dsDNA Dye working solution for a total volume of 200μl in the quantitation tube, then the volume entered on this screen should be 1μl. 9 Measure fluorescence of the unknown sample. Protocols Quantus™ Fluorometer Operating Manual #TM396 » QuantiFluor® dsDNA System Technical Manual #TM346 » Watch a Video on Sample Measurement Using a Quantus™ Fluorometer » enews | July 2015 Pagina 14
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