Chromatography Quick Question

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logisticslord2
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Chromatography Quick Question

Post by logisticslord2 » Wed Apr 04, 2007 5:35 pm

We did chromatography for nucleic acids. There are 3 Rf values calculated. They are .76, .44, and .21. The values given for RNA=.71 and DNA=.39. How do we explain all three of these then? Does it have to do with what bases are present? Help please...if you have a link that's excellent but if not that's cool too..

blcr11
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Post by blcr11 » Wed Apr 04, 2007 7:14 pm

This is paper or thin layer chromatography? And when you say "RNA" and "DNA" are we talking about single nucleotides (base + sugar + at least 1 phosphate), nucleosides (base + sugar but no phosphate) or polymers of same (i.e. oligonucleotides of RNA or DNA)? And then the reagent used to detect the material is general (like a sulfuric acid spray, say) or specific for nitrogens, like a dye or ninhydrin, or UV light which can detect the bases under the correct conditions?

Off hand I would guess you have a mix of nucleosides of RNA and DNA. The material at 0.76 and 0.44 most likely correspond to RNA and DNA. Rfs are notoriously sloppy things to measure--spots are usually large and often mis-shapen which makes it difficult to find the true mobility of the zone. I'm guessing the slow moving material is some free bases that have hydrolyzed off the nucleoside(s). You have to take that interpretation with a grain of salt. I don't know the specifics of your samples, your separation/detection method, or the Rfs of any of the free bases of nucleic acids, so I don't know if that's the correct interpretation or not, but it is my guess.

soso
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Post by soso » Sat Apr 07, 2007 7:41 am

ok..i also wana know dose the RF factor has a standerd for different kind of materials are they a general standers values?? lets say its for Group A from x to y for example and for group B its its from B to C and so on.. are there such a things??

soso
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Post by soso » Sat Apr 07, 2007 7:43 am

ok..i also wana know dose the RF factor has a standerd for different kind of materials are they a general standers values?? lets say its for Group A from x to y for example and for group B its its from B to C and so on.. are there such a things?? not for just nuclic acid but for enzymes and other kind of materials??

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