G-banding mouse spreads

W R Bennett bspwrb at bath.ac.uk
Fri Mar 7 06:33:55 EST 1997


Does anyone know what sort of variables might control whether G-bands are
visible on mouse chromosome spreads?  I ran 4 slides simultaneously
through G-banding yesterday, and one slide shows beautiful banding, and
the rest don't - the chromosomes are evenly stained with Giemsa
throughout.  The spreads were made from different flasks of E15 embryonic
fibroblasts, but were all made on the same day and stored in the same
freezer at -20, and treated exactly the same. 

Banding protocol:  cook slides 90min in 2xSSC at 60C, rinse 3x5min in 
0.9% NaCl, stain 8min in 4% Giemsa, 0.002% trypsin in Gurr phosphate 
buffer pH 7.2 - stain freshly made and filtered, slides washed 4x2min in 
Gurr buffer/water 1:1, air-dried and photographed.

Thanks for any help,

Bill Bennett
Developmental Biology Unit,
School of Biology and Biochemistry
University of Bath

bspwrb at bath.ac.uk



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