Technical Data

1. Spectrum

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Excitation (EX) and Emission Wavelengths

EM scan at 306 nm EX showed the highest center peak at 530-535 nm.

EX scan at 535 nm EM revealed multiple peaks, with the center peaks at 270 nm, 300 nm, 480 nm, and 495 nm.

Excitation Capabilities

RedSafe can be excited not only by the UV light typically generated by a UV Transilluminator, but also by a wide range of visible light wavelengths, specifically in the 420-510 nm range.

Emission Characteristics

Upon excitation, RedSafe emits a strong fluorescent signal at 535 nm, which is in the green-yellow region of the visible spectrum.

2. Filter Selection

The Image Analyzer used after electrophoresis is composed of a UV transilluminator, a camera, and analysis software. In most cases, a filter that can selectively transmit only specific wavelengths of light is either attached or can be selected in front of the camera lens mounted on the equipment.

​When using an EtBr Filter

If the existing laboratory equipment is equipped with an optimal 590 nm filter for EtBr, the fluorescent signal of RedSafe may still be detected, but it may be difficult to obtain the best image.

When using a 520-540 nm range filter

When using a filter in the 520-540 nm range, which is suitable for RedSafe, the RedSafe bands will appear brighter and more distinct. However, in this range, the EtBr bands will hardly be visible. Therefore, if you want to obtain optimal electrophoresis results using RedSafe, it is recommended to use a 530 or 535 nm filter.

When no filter is available

Even if there is no filter suitable for RedSafe, it is still possible to obtain electrophoresis images. However, if the UV transilluminator is set to a high output, the UV ramp behind the gel will be captured, resulting in a blurry background. In this case, reducing the UV ramp output to 40% or less, so that the light bulb is not visible, will allow for good images of both RedSafe and EtBr.​

3. Post-Staining

 

Typically, for easy and convenient electrophoresis, RedSafe is commonly added during agarose gel preparation (Precasting method). However, due to the nature of electrophoresis, as the run time continues, RedSafe tends to migrate upwards (+charge) while DNA moves downwards (-charge). This results in a non-uniform background, where the upper portion appears brighter and the lower portion appears darker (It is a characteristic of all staining dyes, including EtBr).

​Occasionally, there are inquiries about the possibility of Post-staining to maintain a cleaner experimental environment or achieve a more uniform gel background. The good news is that Post-staining is indeed possible.

The procedure for Post-staining is similar to that of EtBr. To perform Post-staining, the final concentration of RedSafe should be diluted to 1x in the running buffer, and the gel should be incubated for approximately 10 minutes. This will result in a clean electrophoresis image with a uniform background. Additionally, the even distribution of RedSafe throughout the gel will allow for relatively bright visualization of even small DNA fragments.