Dear all,
I faced the problem of quantification of input DNA for library preparation process. I attached a gel image showing six different DNA preps. According to nanodrop quantification, they should be (from left): 380ng, 940ng, 1080ng, 1030ng, 638ng, 620ng, marker.
But why there are too much difference between the quantification result of nanodrop and EtBr image.
I always resuspend DNA with RNase for 37C for 1.5h, following with Phenol/chloroform treatment and 2X EtOH (1/10 NaAc) precipitation. DNA looks intact and no RNA contamination appeared through gel image. Thus I don't know what kind of material within my DNA sample made Nanodrop overestimate the quantification.
I always used 5ug DNA as initial input for library. But now it looks I need a totally different way to quantify my DNA or I have some mistakes with DNA extraction?
Looking forward to your help, thanks
SK
I faced the problem of quantification of input DNA for library preparation process. I attached a gel image showing six different DNA preps. According to nanodrop quantification, they should be (from left): 380ng, 940ng, 1080ng, 1030ng, 638ng, 620ng, marker.
But why there are too much difference between the quantification result of nanodrop and EtBr image.
I always resuspend DNA with RNase for 37C for 1.5h, following with Phenol/chloroform treatment and 2X EtOH (1/10 NaAc) precipitation. DNA looks intact and no RNA contamination appeared through gel image. Thus I don't know what kind of material within my DNA sample made Nanodrop overestimate the quantification.
I always used 5ug DNA as initial input for library. But now it looks I need a totally different way to quantify my DNA or I have some mistakes with DNA extraction?
Looking forward to your help, thanks
SK
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