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