High DNA Spec - Nothing on the Gel... - (Oct/24/2008 )

Hi folks,

I am pretty upset by some results I have had lately. I have been using "genomephi" to amplify a small amount of gDNA I have (Starting amount = approx 10ng/ul). After the treatment with genomephi my DNA spec tells me I have up to 900g/ul (in the range of what is expected).

The DNA Spec gives me 2.61 and 1.83 ratios for the DNA I am measuring, and a fastastic Curve. In short nothing to warn me that I have contamination from proteins or other stuff...Everything on my spectro tells, oh my goodness the gDNA you have is fantastic!

However, when I load 7 to 10 (supposedly MICROGRAMS) of this on a 1% agarose gel...I have Nothing...nada...0...the DNA is invisible...

I just dont understand what's wrong.

Any input would be greatly appreciated.

Thanks

This DNA won't run very well on a gel. Is there a bright band at the well? Phi29 produces a tangle of DNA, which will not run unless cut with an enzyme, and then will produce only smears. What do you expect to see on the gel?

I assume you have purified the product, since the dNTPs in the reaction will also give high spec readings, even without long products.