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Trypsinized cells after fixation


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#1 Claudia67

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Posted 10 March 2009 - 04:08 PM

Hi everyone,

Do you know if I can trypsinized the cells after fixation in order to obtain RNAm for RTPCR?

I really appreciate your help.

Thanks in advance.

#2 Mrtn

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Posted 09 August 2009 - 09:00 PM

Do you know if I can trypsinized the cells after fixation in order to obtain RNAm for RTPCR?

Is RNAm a different notation for mRNA (messenger) ?
Why do you fixate cells before extracting (total) RNA?
I've always isolated total RNA from cells with a MN nucleospin column and then used a one- or two step RT kit with oligo dT primers for q-RT PCR.

#3 bob1

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Posted 10 August 2009 - 05:13 PM

Fixing crosslinks the DNA and RNA so that it will be very hard to extract.

#4 DavidJ

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Posted 21 August 2009 - 03:47 AM

It kind of depends on the fixation. I believe you can do it from (snap) frozen cells. Can you try lysing the cell directly. I also do that for non fixed isolations.




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