Take 30µL of 10µM ssDNA (T4 ligation primer) into a new eppendorf

Dilute 10µM ssDNA into 1µM :

Take 3µL of 10µM ssDNA and 27µL RNase-free water into a new eppendorf

Dilute 1µM ssDNA into 100nM :

Take 3µL of 1µM ssDNA and 27µL RNase-free water into a new eppendorf

Dilute 100nM ssDNA into 10nM :

Take 3µLof 100nM ssDNA and 27µL RNase-free water into a new eppendorf

Dilute 10nM ssDNA into 1nM :

Take 3µL of 10nM ssDNA and 27µL RNase-free water into a new eppendorf

Dilute 1nM ssDNA into 100pM :

Take 3µL of 1nM ssDNA and 27µL RNase-free water into a new eppendorf

Dilute 100pM ssDNA into 10pM :

Take 3µL of 100pM ssDNA and 27µL RNase-free water into a new eppendorf

Add 1.5µL of 20X evagree dye into each eppendorf.

Respectively load 20µL of 10µM,1µM, 100nM, 10nM, 1nM, 100pM and 10pM ssDNA into each well

Measure the fluorescence excitation and emission intensity