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Degraded cDNA after freezing


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#1 Mathias Droll

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Posted 06 February 2002 - 12:53 AM

RNA isolation from immunohistochemically stained frozen tissues (Chomczynski) with subsequent RT-PCR works well.
However, after freezing of cDNA the PCR doesn`t work anymore.
There is no cDNA detection anymore.

any suggestions?


(Edited by Mathias Droll at 6:54 pm on Feb. 5, 2001)

(Edited by Mathias Droll at 6:56 pm on Feb. 5, 2001)


#2 Cthulhu

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Posted 08 February 2002 - 07:56 AM

I'm thinking freezing alone could not do this. I sure have never had a problem like this. I freeze my reactions in -20 C. It must be something other than the freezing. Have you solved the problem?

#3 Mathias Droll

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Posted 08 February 2002 - 09:22 AM

hi,
I made another  RNA Isolation + cDNA .
I put one part in the -20C freezer and another part in the 4C Refrigerator.
The next day I made a PCR.
The PCR with the 4C DNA works fine but the PCR with the -20C PCR had no results :(

(Edited by Mathias Droll at 5:12 am on Feb. 9, 2001)





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