From: Ian_DIMMICK@europe.bd.com
Date: Thu Dec 18 2003 - 15:15:23 EST
Dear John, I would recommend the DNA QC particle kit by BD, I must stress
that I am not using this as an opportunity to use this site as a commercial
vehicle, so let me explain why I recommend this kit,
The methodology outlined with the kit is very informative and goes
through the theory of the doublet discrimination procedure
Fluorescent particles are provided within this kit so to evaluate the CV
of measurements on FL2 independent of staining but dependant upon
instrument these particles have a very achievable target value relevant
to DNA analysis
Chicken erythrocytes are provided to ensure linearity on a linear scale
for FL2
Calf Thymocytes are provided with target values for GoG1 / S /G2m this
enables you to visualise also that you have your gate correctly set to
enable doublet discrimination
For steps 3 and 4 the Calf and chicken nuclei are stained with PI
which is included within the kit , I use this kit a lot both for instrument
set up for PI , Hoechst, Dapi and to evaluate and train proceedures for
correct doublet discrimination with DNA analysis.
Ian ,
Wishing everyone a merry Christmas wherever and whoever you are with
"Dr. John Waitumbi" To: Cytometry Mailing List
<cytometry@flowcyt.cyto.purdue.edu>
<JWaitumbi@kisian. cc: mimcom.net> Subject: Re: DNA analysis 18/12/2003 07:55
I'm stuck at the basics. Can someone give me tips on setting up doublet
discriminator on FACScan Lysis II or FACScalibur. I cannot see my PI
stained cells on pulse width or pulse area (set in FL2). I've tried to up
the amplificatiion gain etc but cannot see a single cell on the computer
but can see pulses on the cytometer.
John
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