Could you please guide me about the protocol for conducting electrophoresis at very low concentrations of DNA (below 1 ng/ul)?
I did electrophoresis with two percent agarose gel (+1ul safe stain) and four microliters of DNA + two microliters of loading dye, the DNA concentration was one nanogram per microliter, but I did not see any bands!
If you are running 4ng of dna per well then this is spread over much of the length of the gel (assuming some degradation) and there is just not enough dna to see a signal. You will need to load a lot more dna to se it on a gel or use SYBR Gold stain which is capable of detecting 25pg of dna
Fereshteh Amini 4 ng is too small. try to increase the total DNA you load or you should use another stain method that very sensitive to detect at that low level. Mr Paul Rutland give a very good reccomendation. Also try using a smallest well when making a gel agarose.
Try to increase the total DNA you load and use low concentration of agarose gel with small well. Also use buffer with low Ionic strength and use different dye .
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