After carrying out A260/A280 on extracted DNA has given a value of 1.8 indicating the DNA is pure. Is there other methods to confirm that the DNA is pure?
Qubit is highly recommended/preferred over nanodrop for the DNA analysis for NGS now a days.
https://en.wikipedia.org/wiki/Qubit_fluorometer
https://tools.thermofisher.com/content/sfs/manuals/qubit_3_fluorometer_man.pdf
Bisde what have been suggested, you can run it in a agarose gel and see if you will have a one sharp band
Malee Nagi you can always run an agrose gel as told by Ahmed Dhamad to confirm the purity
Sahithi Nimushakavi and Ahmed Dhamad thank you I was thinking of this but didn't know where an agarose gel or polyacrylamide would work best
Check this link
http://www.mn-net.com/Portals/8/attachments/Redakteure_Bio/Protocols/Genomic%20DNA/UM_gDNABlood_NS8QP.pdf
regards
How pure do you want or need your DNA to be, and why? Can we assume that your gDNA is extracted from ES cells? If so, it will appear as a smear on agarose gels, not a defined band. However, if there is RNA contamination, you will see faster migrating bands of ribosomal RNA subunits (which shouldn't be there, but perhaps your RNase step didnt work effectively). Low level protein contamination (not detected by Nanodrop as the 280max is masked by the DNA 260 peak) is more difficult to quantify, but do you need to know??
You are welcome, I would say you can use either one but polyacrylamide is more efficient to resolve small DNA molecules (5-500bp). Good luck
Ahmed, please explain to Malee (and me) how knowing the size of her DNA fragments tells her how pure they are? And note that if the DNA is from ES cells (according to the header) it will not be a single band, and will probably not move out of the wells of acrylamide gels.
Dear you can use first nanodrop where you observe their Absobance A260/A280 - https://tools.thermofisher.com/content/sfs/manuals/qubit_3_fluorometer_man.pdf
after then with Agarose gel electrophoresis with 1kb marker where you shall be found a sharp band according to their size.
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