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gene expression, reverse transcription on total RNA or mRNA?


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3 replies to this topic

#1 susanna

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Posted 23 December 2009 - 02:02 AM

Hi,

I want to check genexpression. For this purpose, i have totale RNA from my patients. So i wanted to use a reverse transcriptase kit, for cDNA synthesis. Now, i wondered if, i can apply the reverse transcriptase on the total RNA. Or is it suggested, to first isolate the mRNA, from the totale RNA, before doing reverse transcription?
Do anyone has experience with this issue?

greetz,

Susan

#2 mdfenko

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Posted 23 December 2009 - 09:04 AM

the instructions that come with the kit should tell you which it prefers but it should give you the procedure either way.
talent does what it can
genius does what it must
i do what i get paid to do

#3 susanna

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Posted 28 December 2009 - 12:17 AM

the instructions that come with the kit should tell you which it prefers but it should give you the procedure either way.


thats the problem, they are not sure :-/

#4 mdfenko

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Posted 28 December 2009 - 08:38 AM

thats the problem, they are not sure :-/

well, that's unusual. are you sure that they are not just saying that the kit works equally well with either? if so then you may want to avoid additional steps and just use trna.

i prefer to use mrna but i get a large yield of trna (i use a lot of starting material).

just follow the instructions for whichever you choose.
talent does what it can
genius does what it must
i do what i get paid to do




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