We have had a number of requests from users regarding the compatibility of our blue LED transilluminator (470nm) and UV transilluminator (311nm) with popular DNA gel stains. We routinely use GreenView ourselves, but in the past we have also used SYBR® Safe, ethidium bromide, GreenView Plus and RedView. These are summarized in the table at the bottom of the page.
In this blog post we check two more commonly used DNA stains with our transilluminators: GelGreen™ and GelRed™ stains from Biotium. We ordered the 0.1 mL trial sizes for both GelGreen and GelRed (Parts #41005-T and #41003-T respectively). Each stain was used according to the manufacturers recommendations (see Product Information sheets on the website). For testing with our transilluminators, we only used the post-staining method, which we routinely use and prefer. However, these stains can also be added to the agarose gels, the same as with most other gel stains. DNA sample was 100-300ng of a 1Kb DNA ladder (NEB).
Gel Green + Blue LED transilluminator
From the product information sheets, GelGreen has a similar emission/excitation to GreenView - excitation peak around 500nm, emission at 530 nm, so using the blue LED transilluminator we expect a similar result for both DNA stains.
As per our usual method, GreenView was diluted to 2X in TAE buffer (6uL of dye in 30 mL). GelGreen was diluted to 3X in distilled water following the manufacturers suggestions (9uL of dye in 30 mL). After electrophoresis, the gel was divided lengthwise into two gels. Each gel was placed into a plastic container with either GreenView or GelGreen and stained for 30 mins with gentle shaking on a platform shaker.
Although both stains should not require destaining, the GelGreen gel was visibly stained orange/pink (image below left), and we found that we could get better images by destaining the gel for at least 30 mins in water (right image). GreenView gels did not require any destaining step.
Gel Red + UV Transilluminator
From the product information sheets, GelRed has a similar excitation to RedView with an excitation peak around 300nm. The UV transilluminator (311nm) should be suitable for both stains.
As per our usual method, RedView was diluted to 2X in TAE buffer (6uL of dye in 30 mL). GelRed was diluted to 3X in distilled water following the manufacturers suggestions (9uL of dye in 30 mL). After electrophoresis, gels were placed into plastic containers with either RedView or GelRed and stained for 30 mins with gentle shaking on a platform shaker.
No destaining step was required for either stain - both can be imaged on the UV transilluminator immediaetly after staining. The image on the left is taken of the gels without any emission filter, and on the right a Roscolux #19 filter (Rosco) was used to image the gel.
Summary of gel stains tested with transilluminators
This Table summarizes the gel stains that have been tested so far with our two transilluminators. See the images in linked blog posts for examples.