I am using AB Sciex 5800 TOF/TOF MALDI platform for analysis of RNA. For that, initially I am trying to standardize with 20-mer DNA primers by using 3-HPA and THAP as matrix but I couldn't get any spectra. Please help me with it.
Mix "clean" oligonucleotide (~20µM) equal volume with matrix+DAHC solution and spot on target and allow to dry.
It is quite critical that the oligonucleotide be desalted prior to MS analysis as these molecules bind cations readily. I use a Ziptips, dialysis, or centrifugal filtering as buffer exchange techniques to help desalt prior to analysis.
1. You have mentioned three desalting technique. can you suggest me which one is most effective ? can you provide that protocol ?
2. what was the instrument parameter settings ? e.g. - plate voltage, grid voltage ?
3. Right now I am playing with Primers (20 mer, DNA) is it essential to desalt those as the manufacturer mentioned a standard desalting (?) has been done ?
1) I don't work with oligonucleotides all that often, but for proteins, I generally favor desalting using centrifugal filters likes the ones here (http://www.emdmillipore.com/US/en/life-science-research/protein-sample-preparation/protein-concentration/amicon-ultra-centrifugal-filters/YKSb.qB.GXgAAAFBTrllvyxi,nav). If the MW of your target is small (