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RNA Exctraction

About microscopic forms of life, including Bacteria, Archea, protozoans, algae and fungi. Topics relating to viruses, viroids and prions also belong here.

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RNA Exctraction

Postby danaC » Sat Feb 12, 2011 12:16 am

I did an experiment to isolate RNA, we had to centrifuge the E.coli suspensions A and B at 13,200rpm for 2mins. A gave a loose gelatinous pellet while B gave a solid looking pellet. We added digestion solution and incubated it. With A, a white chunky precipitate formed yet B was clear. We lysed ,filtered usin GenElute columns and washed etc. Upon a 1:50 dilution, the ratio of A20/A280 of RNA in A is 1.17 and B we got a ratio of 2. We have to compare and ive reasons for the differences. When i look up a gelatinous pellet, i see that that indicates RNA (http://www.med.yale.edu/mbb/koelle/prot ... _prep.html)...i dont quite understand, any suggestions?
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Postby canalon » Sun Feb 13, 2011 7:10 pm

The RNA gelatinous pellet would be formed after the column treatment, not at the bacterial level. So the question you have to solve is why suspensions A and B are different. There is no link with nucleic acids, and you have to investigate what strains are, and what kind of differences can cause your orginal pellet to misbehave.
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Postby danaC » Sun Feb 13, 2011 9:35 pm

but that is what im trying to figure that out.we are suppose to figure it out ourselves because the gel. pellet makes no sense. we were not given the strains even when asked,they said it was unnecessary.
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