Hi everybody!
I don't know why gel bands appear with low intensity. I use 50 uL SYBR Green I diluted in 200 mL TAE 1X to stain the gel. Also, gel is staining during 1 hour in agitation and darkness. I attach a image.
Some recommendations???
Thanks!
Cristina
Hi Cristina!
What amount of PCR product are you putting in the gels and the concentration of DNA in the PCR products and what denatured gradient did you use? sorry for asking more questions...
Best
Cristina
Hi Cristina,
How are you imaging your gel? UV transilluminator or something else? Check the emission wavelength of whatever you are using to be sure it matches what the stain needs, and that your camera filters are set correctly for the stain emission wavelength. I also found bands were weak when using UV, so I switched to using a Typhoon which matched my stain requirements much better. Otherwise, I would do as Cristina suggested above and check your DNA concentrations: you may just need to add more per well.
It looks like you've got nice clear bands so once you've increased the intensity they should be good to work with!
Good luck!
Alison
Thank you very much!!! Denaturing gradient is 35-65% and the amount of PCR products are 8 uL with 7 uL of loading buffer 2X. Concentration of DNA is around 70 ng/uL
Hi Cristina
I think the problem of your gel is linked to the concentration of DNA in your PCR product.
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