[Yeast] FACS sorting/Regeneration of spheroplast

Wang, Xin-Xiang via yeast%40net.bio.net (by WangXX At MedImmune.com)
Tue Jan 30 17:27:11 EST 2007


Sure.  But we are interested in something in the periplasmic space.  We
seeminly cannot partially disrupt the cell wall while accessing the
interested product.  We have to make spheroplast in order to detect it.
Thanks!  

My question is: can the spheroplast (do delicate after removing the cell
wall) sustain the electro-charging during FACS sorting? I know you can
do flow cytometric testing with spheroplast, though.  Any experience?  

Xinxiang

-----Original Message-----
From: Eric Grote [mailto:egrote At jhsph.edu]
Sent: Tuesday, January 30, 2007 5:11 PM
To: Wang, Xin-Xiang
Subject: Re: [Yeast] FACS sorting/Regeneration of spheroplast


If you reduce the spheroplasting time, they will regenerate better.


On 1/29/07 12:45 PM, "Wang, Xin-Xiang" <WangXX At MedImmune.com> wrote:

> Hello,
> 
> Has anybody done FACS sorting with spheroplast and then regenerate
them in Top
> agar?  Would the charge in FACS machine burst the spheroplast during
sorting?
> My regeneration rate was only 1%.  Any hint to improve that? Thanks!
> 
> Xinxiang
> 
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