dsDNA quantification calculator

Use this calculator for fast and easy quantification of dsDNA with our Pico488 DNA quantification kit. The linear regression tool can be used for any type of quantification. The dilution series and recommendations also fit the most popular format of dsDNA quantification reagents: the Quant-iT® PicoGreen® dsDNA quantitation assay kit.

Fluorescence - background, a.u.

Concentration, pg/µL

Cuvette volume: µL
Cuvette volume should be at least 1 µL.

1. Preparation of buffer and Pico488 working solutions

• Dilute 592 µL of the 20× TE buffer stock solution (part of the kit) with 11.248 mL of deionized water to obtain of 1× ТЕ buffer.
• After thawing, dilute of the Pico488 dye stock solution (part of the kit) 200-fold with of 1× ТЕ buffer. (Pico488 is also available separately as quantification reagent.)
• Mix and use up within the next 3 hours.

2. dsDNA standard serial dilution

• MixTake of a ng/µL dsDNA standard solution Concentration is too low! Should be at least 2 ng/µL. (a 100 ng/µL solution is part of the kit) with of 1× TE buffer to obtain a dsDNA stock solution of 2 ng/µL.
• Further dilute this dsDNA stock solution with 1× TE buffer and add the Pico488 working solution according to the following table.

  #	1× ТЕ buffer, µL	dsDNA stock solution	Pico488 working solution, µL	Final dsDNA concentration, pg/µL

3. dsDNA sample dilution and fluorescence measurements

• Add µL of your DNA sample to of 1× TE buffer, then add of the Pico488 working solution. Mix and incubate for 5 min.
• Measure the fluorescence intensity of standard and sample DNA solutions in a fluorometer. Excitation maximum is at 503 nm, emission maximum at 525 nm.

  #	Concentration, pg/µL		Fluorescence intensity, a.u.	Calibration chart
  	Calibration data
  #1
  	DNA Sample	DNA sample dilution