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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.

Bestandteile

Komponente Anzahl
12102
100 assays
13102
100 assays (incl. tubes)
52102
500 assays
dünnwandiges transparentes 0,5 mL Polypropylen-Gefäß 100
63010, QuDye dsDNA HS Reagent, 1.25 mL 1
G9650, dsDNA quantitative standard, 0 ng/uL in TE buffer, 5 mL 1
G7650, dsDNA quantitative standard, 10 ng/uL in TE buffer, 5 mL 1
N2150, TE buffer, 20x, 25 mL 1
33010, QuDye dsDNA HS Reagent, 250 uL 1 1
B9650, dsDNA quantitative standard, 0 ng/uL in TE buffer, 1 mL 1 1
B7650, dsDNA quantitative standard, 10 ng/uL in TE buffer, 1 mL 1 1
G2150, TE buffer, 20x, 5 mL 1 1

bei +4 °С lagern

Haltbarkeit: 12 Monate

Protocol

  1. Prepare 1× TE buffer by diluting the 20× TE concentrate 1:20 with deionized water.
  2. 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.
  3. 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.)
  4. 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.
    Vortex all tubes for 2–3 seconds and incubate them for 3–5 minutes at room temperature. Conduct the fluorescence measurement according to the manual of your Qubit® instrument.

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.

  1. On the home screen of the Qubit® fluorometer select the assay type "dsDNA HS (DNA High Sensitivity)" and press "Go".
  2. With every new preparation of the QuDye working solution, the fluorometer must be calibrated. 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. 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 volume

    The 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.

Related kits

QuDye dsDNA HS Kit (optimiert für Qubit®)

Für Qubit®-Fluorimeter optimiertes Kit zur Quantifizierung doppelsträngiger DNA mit hoher Sensitivität (HS) und einem Messbereich von 10 pg/µl bis 100 ng/µl
Preise einblenden
Artikel-Nr. Packungseinheit Preis Vorlaufzeit
12102 100 assays $55.00 Auf Lager
13102 100 assays (incl. tubes) $70.00 Auf Lager
52102 500 assays $255.00 Auf Lager
53102 500 assays (incl. tubes) $299.00 Auf Lager

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