[Drosophila] RNA and RT-PCR

Kathleen Gajewski via dros%40net.bio.net (by sa08383 from wotan.mdacc.tmc.edu)
Thu Jul 12 11:05:15 EST 2007


Dear Arindam,
	I use TRIzol to prep my RNA.  I haven't tried embryos, but I have had 
success with RT-PCR using whole adults and larvae, IFM dissected from 
freeze dried adults, and even pooled blood samples from 3rd instar 
larvae.  I'll bet it would work well for embryos too.  I also use the 
Invitrogen kit.

KG


On Jul 11, 2007, at 7:47 PM, Dr. Arindam Basu wrote:

> Dear All,
>    I am having problems with getting good cDNA synthesis with 
> Drosophila total
> RNA harvested from embryos. Quite a few times the actin PCR does not 
> show a
> good signal for checking the RT reaction. Are there any special 
> step(s) for RNA
> extraction and/or Reverse Transcription reaction (I use Invitrogen 
> Superscript
> kit for RT) to get optimum cDNA synthesis?
>
> Thanking you in advance
> Arindam
>
> -- 
>
>
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