How much RNA in 24 hpf Zf embryos?

Adam Rodaway adam.rodaway at kcl.ac.uk
Fri Nov 15 14:41:41 EST 2002


Hi Corinne,

for RT-PCR, 50 embryos should give you plenty of cDNA.

I think  this many embryos will probably contain about 10ug of total 
RNA, with around 1-300ng being mRNA. I've recently used these 
estimates for doing RACE, and it's worked well. I used the 
'Oligotex Direct mRNA mini kit' from Qiagen to directly isolate 
mRNA for use with Invitrogen's GeneRacer RACE kit.  

I've previously used similar numbers of embryos to do semi-
quantitative RT-PCR (I used random 10-mers and AMV RT on total 
RNA) and found I had enough cDNA to last a lifetime!  

hope this helps,
Adam Rodaway

Corinne Renier wrote:

>Hello,
>
>I need to prepare total RNA from embryos as early as  24 hpf and I
>was wondering how many embryos I would need to get enough material to
>play with (RT-PCR). Does anyone of you could share his/her experience
>with me? Thanks a lot. Corinne  
>
>----
>

-- 

Dr Adam Rodaway
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