Extarction and purificaton of DNA and RNA

[G Campbell]

Extarction and purificaton of DNA and RNA

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Post by G Campbell » Tue May 09, 2006 1:30 pm

Could someone give me sites the explains it because my exam in molecular biology is next monday and this is a possible exam question? I have looked in google but I dont really get anything usefull.

Thanks in advance

[LilKim]

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Post by LilKim » Tue May 09, 2006 2:20 pm

Hey G Campbell....

Can't think of any sites off-hand. And there are several protocols that effectively extract nucleic acids ...(they're slightly different)

But, if you tell us what you know about DNA or RNA extraction those of us who have done it (a MILLION times) may be able to help clarify the points of the protocol.

good luck!
- KIM

[canalon]

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Post by canalon » Tue May 09, 2006 3:06 pm

In short:
-break the cells
-get rid of cell debris
-Degrade DNA or RNA (if you want the other)
-Clean DNA and/or RNA of proteins
-concentrate and wash all unwanted reagents from the previous reactions
-Resuspend in appropriate buffer.
A million protocols using 2 few basic techniques to clean the DNA/RNA (lysis just depends on your starting material) ie Phenol chloroform and Silica/NA interactions exist.

You can try to search http://www.protocol-online.org for detailed protocols.

[G Campbell]

Making a gene

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Post by G Campbell » Sat May 13, 2006 5:35 pm

Are all these the right steps in making a gene and am I lacking in information? If you have the time to read it quickly and give feedback it would be appreciated.

[canalon]

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Post by canalon » Sun May 14, 2006 1:31 am

something is missing here....

[G Campbell]

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Post by G Campbell » Sun May 14, 2006 2:31 pm

I can't seem to attach it but are these all the steps needed in making a gene:

1. Digest DNA with restriction endonucleases
2. Gel electrophoresis
3. With the fragments use the southern blot method
4. Add a probe
5. Use the PCR method

Is that right?

[canalon]

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Post by canalon » Sun May 14, 2006 4:45 pm

OK. I see we have a problem here. What do you call "making" a gene.

But whatever you want to do, your steps do not really make sense to me. A short list of problems:
1/ what DNA?
2/Of what, it's OK if it's 1, but depends on which DNA you cut in the first place.
3/What probe? to detect what?
4/The probe is part of the Southern procedure. In fact a Southern is the use of a probe it doesn't make sense
5/To amplify what, from which template? with which primers? And why is that coming after the Southern?