I use 2% Agarose gel with 90V and 60 minutes for my PCR product. I'm confused why my 100bp DNA ladder not separates well in SYBR Green but separate well enough when I'm using SYBR Safe with the same percentage of agarose gel. I've tried to change the voltage and adjust the time when I run it but the result is still the same. Is it because of the concentration of the SYBR Green 10,000X with DMSO or I need to adjust another thing?
Hello Norhidayah, how are you? I’m not sure about differences between SYBR Green and SYBR safe, if I am not mistaken, they are the same dye, and you would not have problem. However, the SYBR green it is not made for reveal DNA or cDNA in agarose gel. SYBR safe is the correctly dye for that. If you are adding SYBR green when you preparing the gel this will probably affect the running due the differences in formulations. SYBR green is suitable for qRT-PCR in real time machines. I never use SYBR green for agarose gel and I not recommended you do that.
I recommend using SYBR Safe or GelRed only for agarose electrophoresis.
However, if you are really curious, you could try staining your gel with SYBR Green AFTER running your DNA samples. To do so, simply dilute your stain 1:10 000 in your running buffer, and soak your gel into that solution for about 40mins, protected from light. If you get a much cleaner ladder this way, it means the DMSO from SYBR Green affects the migration of your samples.
Hi Tulio Teruo Yoshinaga I'm fine thank you. How are you? Thank you for the information about the application of the SYBR Green because I only use GelRed and EtBR for my previous project.
Hi Amy. Thank you for the suggestion.
Hi Martin. I will try it using your suggestion. Thank you.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- DNA