dsGreen is a less toxic alternative to Ethidium bromide. The dsGreen dye is a highly sensitive stain used for the visualization of dsDNA in agarose and polyacrylamide gels.
dsGreen has a number of advantages compared to routinely used Ethidium bromide:
✔ 25-fold more sensitive for dsDNA and 50-fold for RNA detection
✔ much less mutagenic
✔ can be used with either blue light or UV excitation
Among others, Gel Imaging Systems supplied by Bio-Rad are suitable for the visualization of gels stained with dsGreen. The image of the gel stained with dsGreen should be taken using
· the XcitaBlue Conversion Screen for Gel Doc™ XR+ System
· UV Tray or Blue Tray for Gel Doc™ EZ System.
As a first step gel should be stained with dsGreen. There are three methods to stain a gel with dsGreen, but we recommend the gel soaking method, which has been shown to be the best method in terms of the separation quality of the DNA fragments and the analysis sensitivity. After the gel has been stained with dsGreen, it will take you a few minutes to visualize the gel.
Gel Doc™ EZ System offers many options for advanced users such as creating default protocols, analysis, quantity tools, etc. In this article, we demonstrate a way to simply take an image of the gel you have stained with dsGreen. For detailed information about Bio-Rad's Gel Imaging Systems, refer to the manufacturer's documentation.
Step 1.
Turn on the imager and start the Image Lab software.
After the front panel LEDs have stopped flashing, the ‘ready’ LED should light up green.
Step 2.
Place a gel on the UV Tray or Blue Tray and insert it into the imager. Close the door.
For visualization of gels stained with dsGreen you can use both the UV Tray and Blue Tray.
Step 3.
On the toolbar, click New.
*If you have created a default protocol, simply press the green Run button on the front of the imager. The default protocol runs automatically.
In the Gel Imaging pane, click Select and choose Nucleic Acid Gels -> SYBR Green
As an Image Color, select SYBR Green.
You can view the image with any color scheme, but SYBR Green color makes viewing natural.
Click Run Protocol. You will see the imaging run progress bar.
A new window will appear, and you will see the image of your sample gel.
In this window, you can process the image.
To export the image, click File -> Export -> Export for Publication.
Select the resolution and dimensions. Click Export and save the image in the folder you wish.
SYBR® Green I is a trademark of Thermo Fisher Scientific
Gel Doc™ is a trademark of Bio-Rad Laboratories