Re: Plant ploidy

Hi Sue:
One of my investigators used the protocol on this website 
http://www.ueb.cas.cz/Olomouc1/LabDol/Research/Flow_cytometry/Protocols/Two-step_proc.htm and had very nice histograms with PI. Have your investigator give it a try.
Joanne Lannigan

On Tue, 11 Mar 2008 17:14:41 +0000
  S Newton <S.Newton@sheffield.ac.uk> wrote:
> Hello all
> 
> I was hoping for a few words of wisdom regarding looking at plant ploidy 
>by
> flow. I have attached the protocol our user is following, when the samples 
>come
> to me, I cannot get a clean peak. There is so much debris in there that 
>any G1
> peak is either very broad, or masked by the debris, it is also very 
>difficult
> to determine if there are any subsequent peaks. I am using either a 
>FACSCalibur
> or FACSort to look at these samples, and the DNA is stained with PI.
> 
> I have tried thresholding on FSC then on SSC then on FL3 and finally on 
>FSC and
>FL3, but still no joy.
> 
> Any advice greatly appreciated (as ever)
> 
> Sue
> -- 
> Sue Newton
>Flow Cytometry Technician
> DU27. D Floor
> Medical School
> University of Sheffield
> Beech Hill Road
> Sheffield
> S10 2RX
> 
> Phone number: 0114 271 3023