I need to run an agarose gel that has restricted DNA however I need to know the minimum amount of dna that can be seen on the agarose gel. I presently do not get visible clear bands due to the concentration.
The minimum amount visible using EtBr is 10ng per band or even less (though I never saw anything below 5ng per band). The concentration of the gel is mainly important for the DNA separation - the bigger your DNA is, the lower is the percentage of the gel. How big is the DNA you want to seperate?
It also depend on the number of cycles you used. If you are using 30 cycles you need like 5-10 micro-liters from your DNA if you are using like 4-6 cycles then you need more. I hope this answers your question.
With EtBr you should be able to see your product starting with DNA concentration of about 20 ng. After standard PCR I use 2 micro-liters mixed with loading dye and water to detect if product is present. (if reaction is standardized before). If you presently do not get visible bands, you should improve PCR.
The minimum amount visible using EtBr is 10ng per band or even less (though I never saw anything below 5ng per band). The concentration of the gel is mainly important for the DNA separation - the bigger your DNA is, the lower is the percentage of the gel. How big is the DNA you want to seperate?
I isolated DNA from leaves by DNA isolation kit, but DNA concentration level is very low 4-50 ng what would be the reason. please suggest
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