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RT problem !!


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#1 mybioweb2

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Posted 04 October 2004 - 02:38 PM

Hi everyone

Has somebody any problem with RT reaction when using RNA extracted with RNeasy Plant mini kit as matrix for RT enzyme? Would kit components contain some inhibitors which minimize the efficincy of RT?

any help would be appreciated

#2 tuckern

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Posted 06 October 2004 - 05:22 AM

The kit components should not interfere with any downstream applications such as RT reactions. It is more likely that your target is not expressed very much (if at all) or that your RNA is knackered/contaminated. I use ordinary RNeasy kits with E. coli RNA and it seems fine. An internal control (in my case i use 16S RNA) of some gene that you know should be expressed is probably a good idea.

What is the context of your RT reaction? Primer extension? RT-PCR?




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