1 x TE working solution was prepared by diluting the concentrate 1:20 x in sterile, distilled DNase free water.
Use this 1 x TE buffer as blank and for preparing all the DNA standard dilutions.
---PicoGreen working solution
Dilute the PicoGreen Reagent 1:200 in 1 x TE buffer in a plastic tube (protect from light). For the 96-well plate 100 ul of diluted reagent is needed/well
Note: This solution must be used within a few hours after preparation.
---DNA Standard Curves
1. Dilute the provided Lambda DNA standard 1:25 in 1 x TE buffer (4ug/ml).
2. For the standard curve, prepare further dilutions of this 4ug/ml DNA stock in 1 x TE buffer =2025ng/ml, 675ng/ml, 225ng/ml, 75ng/ml and 25ng/ml. Use TE buffer as a blank.
Procedure
l Pipette 100 ul blank, DNA standard dilutions or samples to the wells of a white or black 96-well strip plate.
l Add 100 ul of the diluted PicoGreen reagent.
l Mix well and allow standing at room temperature for 2-5 min protected from light.
l Measure with fluorescent microplate reader. Set the wavelengths of the filters used are excitation 485 nm and emission 538 nm or 518 nm.
l Generate standard curve. Read the unknown samples from the curve.
