QuDye dsDNA HS Assay Kit manual

The kit is used for quantification of dsDNA with Qubit® Fluorometer. All reagents are included and optimised to fit Qubit® requirements and measurements range. Kits with x3102 part number include thin-walled transparrent tubes fitting Qubit® instrument.

Kit components

ItemQuantity
QuDye dsDNA HS reagent, 250 uL1 ea
dsDNA quantitative standard (10 ng/uL), 1 mL1 ea
TE buffer 20x, 5 mL1 ea
0.5 mL thin-walled transparrent polypropylene tubes (for x3102 kits only)100 ea

Store at temperature +4°С.

Shelf life 1 year.

Protocol

  1. Prepare the 1x TE buffer by diluting the 20x TE concentrate 1:20 by deionized water.
  2. Prepare the QuDye working solution by diluting the QuDye HS reagent 1:200 with 1x TE buffer (you need 200 μL of dye working solution for each standard and sample).
  3. Set up two 0.5 ml tubes (thin-walled and optical-transparent tube) for the standards and one tube for each sample. Tubes are included in x3102 kits only.
  4. Prepare standards and samples for concentration measurement.
    • To prepare Standard #1 mix 190 uL of QuDye HS working solution and 10 uL of 1x TE buffer in 0.5 ml tube.
    • To prepare Standard #2 mix 190 uL of QuDye HS working solution and 10 uL of dsDNA quantitative standard in 0.5 ml tube.
    • To prepare Samples mix 180-199 uL of QuDye HS working solution and 20-1 uL of user DNA sample (the total volume should be 200 uL) in 0.5 ml tube.
    • Vortex all tubes for 2-3 seconds and incubate the tubes for 3-5 minutes at room temperature and perform measurements according to the manual to your version of Qubit® instrument.

Measurement procedure in Qubit®

The next steps should be carried out according to the instructions of the fluorometer. Depending on the version of the fluorometer the menu items may differ from the specified below.

  1. On the Home screen of the Qubit fluorometer, you should choose "dsDNA HS (DNA High Sensitivity)" as the assay type. Press "Go"
  2. With each preparation of the working solution, you must calibrate the fluorometer. Select "Run new calibration" and press "Go".
  3. 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.
  4. Insert the tube containing User sample into the sample chamber, close the lid, then press "Go". On the screen you will see the QF value. Calculate the concentration of the sample by the formula: Concentration of the sample = QF value x 200/sample volume.

Related kits

QuDye dsDNA HS Assay kit (optimized for Qubit®)

Ready kit optimised for Qubit® fluorometers featuring high sensitivity (HS) and a measurements range from 10 pg/uL to 100 ng/uL
Show pricing
Cat. # Quantity Price Lead time
12102 100 assays $55.00 in stock
13102 100 assays (incl. tubes) $70.00 in stock

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