I read that after HinfI and RsaI digestion the non-telomeric fragments are typically
You can use more than two enzymes like MspI. You need to use teomere specific probe to determine telomere specific fractions. Technique conditions are same for any organism only you have to use the probe accordingly.
Thanks Anukana, I'll consider adding in another enzyme! I'm assuming this is still not completely effective at digesting the DNA; otherwise anything over 1 kb on the gel could be called telomere without doing the southern blot. I'm curious as to what proportion of the > 1kb fragments are telomere vs gDNA. This seems like a basic question to me but I'm not seeing it discussed anywhere, so I must be missing something.
There are regions in gDNA which would not have that many cut sites so gDNA can go upto 1 kb. But the main reason you need to do southern blot, is because the low conc of telomeric DNA is not visible by ETBr.
That's interesting; could telomeres be size-sorted using the BluePippin? Size selection is performed according to a labelled DNA marker; you could get a concentration reading for each size-selection aliquot.
I don't think so. You will get a mix of telomeric and genomic DNA around 1kb. Plus if you size sort the DNA, you are actually missing the point of measuring average telomere length.
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