Hi there!
This week I extracted DNA from leaves using the protocol for HMW DNA, which employs Carlson lysis buffer + Genomic Tip columns (Qiagen). I quantified it with Qubit (920 ng/ul) and Nanodrop (2.500 ng/ul; 260/230: 1.85; 260/280: 2.05) and then I runned an 1% agarose gel electrophoresis (1 hour, 100 volts) and this is what I got:
My sample is in lane 2.
Never seen before a DNA migration like that.
Not sure if this is actually how HMW DNA should migrate under running conditions as described above.
I really appreciate your thoughts about this.
Cheers,
Milton
I think this looks fine!
It looks like a smear when the DNA-"Fragments" are too big to enter the gel and when the DNA is not yet completely dissolved.
If you run a 08,% Agarose-gel the DNA should be able to run into it but would also build a smear.
And your ratios are also good.
I agree with Fabienne Pritsch, the DNA is probably fine, although the smear below the main band suggests that there is some degradation generating smaller fragments. Looking at the full picture, the main band does enter the gel, with little material remaining in the loading well, so poor dissolution doesn't seem to be an issue
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