Help with mRNA purification and DNA polymerase!

[goose_keeper]

Goodday ,

Here are questions extracted from my exam paper, and I don't know
how to anwser perfectly. So anyone interested in the questions ,help
me please!

(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!

[Morris]

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.

[sdekivit]

Goodday ,

Here are questions extracted from my exam paper, and I don't know
how to anwser perfectly. So anyone interested in the questions ,help
me please!

(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!

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.

[goose_keeper]

Thanks for your replies !

Morris said,

I have a protocols about RNA extraction, but they're in Italian... If you're interested i can traslate in English.

How to find your protocols since I can't speak Italian language?

To sedkevit
The Affinity chromatography is really an efective method for mRNA.

[Morris]

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.

[canalon]

How to find your protocols since I can't speak Italian language?

You will find all you need here:http://www.protocol-online.org/

PAtrick

[Jelanen]

Whoa..thats awesome...

-Jel

[VeeKAY]

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.

How to find your protocols since I can't speak Italian language?

You will find all you need here:http://www.protocol-online.org/

PAtrick

[blcr11]

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.