Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.
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Here are questions extracted from my exam paper, and I don't know
how to anwser perfectly. So anyone interested in the questions ,help
(1) Design a method to purify the mature mRNA in the eucaryotic cell.
(2) DNA polymerase has proofreading function, while RNA polymerase cannot proofread the wrong nucleotides. Explain the difference on the basis of biology.
LOOKING FORWARD YOUR REPLIES URGENTLY!
I have a protocols about RNA extraction, but they're in Italian... If you're interested i can traslate in English.
I think that RNA polymerase hasn't proofreasding activity because, I think, RNA damaged or with wrong nucleotides is degradated by Rnases, but in DNA replication an error can bring dangerous mutations, that can be transferred to daughter cell. I think for this is important that DNA polymerse can correct immediately the wong nucelotide.
the purification of mRNA can be done with affinity chromatography at high salt concentration, by using a poly dT group where mRNA can bindt to with it's polyA-tail.
The fact that RNA-polymerase doesn't have proofreading is that a mutation in RNA is degraded by the ubiquitin pathway and is just for 1 protein, whil a mutation in DNA is forever.
Thanks for your replies !
How to find your protocols since I can't speak Italian language?
The Affinity chromatography is really an efective method for mRNA.
My protocols isn'ti n internet because comes foem an experiment, but I can pass it tp you by pm... When I have traslated it.
'It is futile to pretend to the public that we understand how an amoeba evolved into a man, when we cannot tell our students how a human egg produces a skin cell or a brain cell!'
Dr Jérôme J. Lejeune
I HAVE BEEN TRYING TO PURIFY MY GST-FUSION PROTEIN USING THE GLTATHIONE AGAROSE COLUMN AND AFTER RUNNING MY FRACTION SAMPLES ON SDS PAGE I ALWAYS SEE THREE BANDS INSTEAD OF ONE BAND ON MY ELUTED FRACTION. CAN YOU PLEASE HELP ME SO AS TO PURIFY MY FUSION PROTEIN AS A SINGLE BAND.
Invitrogen links to protocols-online. My country portal is the US so I get everything in English, but maybe if you enter through the Italian port to Invitrogen you get the protocols in Italian? Don't know.
9 posts • Page 1 of 1
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