Re: sorting specific chromosomes

Hi Amanda
Almost all human chromosomes can be distinguished from one another
using the base pair biased DNA dyes Hoechst 33258 and Chromomycin A3. To
do this you need a sorter with two water-cooled argon lasers, one tuned
to multiline uv and the other to 457.9nm. You could separate chromosome
8 quite well with just uv. You might be able to separate it with PI or
ethidium bromide or some other dyes but the purity wouldn't be that
great. You also need your cells to grow in culture nicely. Various Flow
Cytometry books have protocols for preparing chromosomes written by
myself and others. Practical Flow Cytometry Edited by Mike Ormerod is
one.
Its all quite involved if you just want to do it one or twice I'd try
get some one else to do it for you.
Its quite fun to do but takes a bit of practice to get decent
chromosome preps and the instrument has to be aligned really well.
If you need I can probably dig out a book chapter and send it to you.
Best of luck

Simon Monard
Trudeau Institute
154 Algonqin Ave
Saranac Lake
New York
12983
Ph 518 891 3080


>>> "Amanda Reichert" <amandare@cancerboard.ab.ca> 03/09/06 7:24 pm >>>

Hi!
I have a client interested in sorting human chromosomes, specifically
chromosome 8.  I imagine that I need to find some way to label
chromosome 8... any suggestions? Also, does anyone know of any protocols
for sorting specific chromosomes or know where would be best to find
flow cytometry protocols? 
Thanks, 
Amanda.
Amanda Reichert
Research Technologist
Flow Cytometry Facility
Cross Cancer Institute
11560 University Avenue
Edmonton, Alberta
T6G 1Z2


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