Hello all,
I have taken the 1D proton NMR spectrum of my DNA sample in 90% H2O and 10 % D2O. I have also taken the spectrum of my sample with the ligand (dissolved in deuterated acetonitrile) under the same conditions.
To identify the change chemical shifts, can I use the D2O peak at 4.7 ppm to align all the spectra? If not, why?
As a workaround, I am thinking of adding 50 uM DSS to each sample and repeating the experiment and then aligning the peaks using the DSS peak. I would like to keep this as a last resort, though.
Thanks,
Subramaniyam
Yep, @Michael is correct. We have performed a lot of titrations. If you are lucky, there will be little overlap and you do not need to add much DSS - just enough to see the resonance clearly in your spectrum. Too much (in relation to your protein signals) of course will cause you to loose S/N by having to record at lower receiver gain.
Both Michael and Kurt gave you the correct answer. HDO peak position is too variable to "align" your spectra. Adding DSS is the way to go.
Thank you all for your replies. I am performing the experiment again with 50 uM DSS added to the sample.
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