Better images of agarose gel - My bands are not clear when i take a image for processing (Dec/29/2010 )
Hi All,
I am facing a problem related with my gel image processing.
When i observe my gel under a gel documentation i can see clear bands at the region of interest. but when i try to take the image they become faint. It has been really a big trouble for me because i need to post those images for my work.
I tried using Photoshop to brighten my image bands, but it didn't help.
Hope anyone would suggest me either How to increase the intensity of my bands or how to give better resolution for my images.
Thnak You in advance
Portofino on Thu Dec 30 03:39:39 2010 said:
When i observe my gel under a gel documentation i can see clear bands at the region of interest. but when i try to take the image they become faint. It has been really a big trouble for me because i need to post those images for my work.
Hope anyone would suggest me either How to increase the intensity of my bands or how to give better resolution for my images.
From a similar problem I had faced, I am assuming that you stain your gel that after electrophoresis, so thats with diluted EtBr. Considering this is correct, I would suggest, staining for a little while longer,,, another five minutes say... and taking the picture a little sooner.... (This is because I had observed that the longer you expose your gel to UV-light, the fainter the bands get... )... if not this, then I am clueless.... and forgive my blabbering....
gt_ameya on Thu Dec 30 05:40:49 2010 said:
Portofino on Thu Dec 30 03:39:39 2010 said:
When i observe my gel under a gel documentation i can see clear bands at the region of interest. but when i try to take the image they become faint. It has been really a big trouble for me because i need to post those images for my work.
Hope anyone would suggest me either How to increase the intensity of my bands or how to give better resolution for my images.
From a similar problem I had faced, I am assuming that you stain your gel that after electrophoresis, so thats with diluted EtBr. Considering this is correct, I would suggest, staining for a little while longer,,, another five minutes say... and taking the picture a little sooner.... (This is because I had observed that the longer you expose your gel to UV-light, the fainter the bands get... )... if not this, then I am clueless.... and forgive my blabbering....
also, you can prolong the exposition on your camera for 2'' and play a bit with the contrast. that usually helps me a bit.
Be sure to destain (in distilled water for ~15 minutes) as well. I don't know whether you're adding the EtBr to the gel before running or submerging the gel in an EtBr solution after running, but if you're submerging the gel in EtBr solution after running, destaining is critical to increase the band contrast. I don't know if destaining helps when you add EtBr to the gel as I never do it that way -- whichever way you're doing it, you might try the other way to see if your results improve.
Also, limit the time that the gel is exposed to UV, as the EtBr florescence bleaches out.