QuDye dsDNA HS Assay Kit manual
This kit is used for the quantification of double-stranded DNA with a Qubit® fluorometer. All reagents are included and optimized to match the fluorometer’s specifications and measurement range. Kits with part number x3102 also include thin-walled transparent tubes fitting the Qubit® instrument.
|QuDye dsDNA HS reagent, 250 µL||1 ea|
|dsDNA quantitative standard (10 ng/µL), 1 mL||1 ea|
|TE buffer 20×, 5 mL||1 ea|
|0.5 mL thin-walled transparent polypropylene tubes (x3102 kits only)||100 ea|
Store at +4°С.
Shelf life: 1 year
- Prepare 1× TE buffer by diluting the 20× TE concentrate 1:20 with deionized water.
- Prepare a QuDye working solution by diluting the QuDye HS reagent 1:200 with 1× TE buffer. 200 µL of the QuDye working solution are needed for each standard and sample.
- Set up two thin-walled and optically clear 0.5 ml tubes for the standards and one tube for each sample. (Tubes are included in kits with part number x3102 only.)
- Prepare the standards and sample dilutions:
- For Standard #1 mix 10 µL of 1× TE buffer and 190 µL of the QuDye HS working solution in a 0.5 ml tube.
- For Standard #2 mix 10 µL of the dsDNA quantitative standard and 190 µL of the QuDye HS working solution in a 0.5 ml tube.
- Dilute 1–20 µL of each DNA sample with 199–180 µL of the QuDye HS working solution in a 0.5 ml tube to a total volume of 200 µL.
Fluorescence measurement with a Qubit® fluorometer
The next steps should be carried out according to the instructions of the fluorometer you are using. Depending on the instrument version, the menu items may differ from the ones specified below.
- On the home screen of the Qubit® fluorometer select the assay type "dsDNA HS (DNA High Sensitivity)" and press "Go".
- With every new preparation of the QuDye working solution, the fluorometer must be calibrated. Select "Run new calibration" and press "Go".
- Insert the tube containing Standard #1 into the sample chamber, close the lid, then press "Go". When the reading is complete (~3 seconds), remove Standard #1. Insert the tube containing Standard #2 into the sample chamber, close the lid, then press"Go". When the reading is complete, remove Standard #2.
- Subsequently insert the tubes with DNA samples into the sample chamber, close the lid, then press "Go". The QF value displayed on the screen is used to calculate the dsDNA concentration of the sample with the following formula:
DNA Concentration = QF value × 200 ÷ sample volumeThe sample volume is the number of microliters you added to the QuDye HS working solution. The result has the same unit as the QF value.
|12102||100 assays||$55.00||Auf Lager|
|13102||100 assays (incl. tubes)||$70.00||Auf Lager|
|52102||500 assays||$255.00||Auf Lager|