HydraGreen™, 20,000X in water, is a non-carcinogenic and non-toxic alternative to Ethidium bromide used for the
detection of nucleic acids in agarose gels. It is as sensitive as Ethidium bromide. There is also no toxic DMSO as HydraGreen™ is supplied in water.
HydraGreen™ has fluorescence excitation maxima at 295 nm and 490 nm. The fluorescence emission maxima is similar
to EtBr when bound to DNA – at 530 nm.
Protocol:
Note: It is highly recommended agarose gel to be ≤0.5cm in thickness. TAE buffer system is preferred for higher
sensitivity.
PRE-STAIN :
Wait for melted agarose to be cooled to 55ºC.
Add 4-6 µl per 100 ml of agarose or 2-3 µl per 50 ml of agarose, swirl and mix well.
IMPORTANT : HydraGreen ™ is supplied in 20,000X concentration in water
Or
POST-STAIN :
Use 10 to 15 µl per 100 ml of staining solution. Same as when using ethidium bromide. For an average gel thickness of
about 0.5 cm, stain 30 minutes, followed by a destain of 30 minutes in water. Protect gel and staining solution from
light with aluminum foil or place in dark.
DETECTION :
Detect bands under UV illumination (yellow or green gelatin- or cellophane filters is recommended for clearer bands) or
non-UV LED illuminators such as Blue Light LED illumination.