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International Journal of Cell Cloning, Vol 3, 133-142, Copyright © 1985 by AlphaMed Press

ORIGINAL ARTICLES

A micromethod for chromosome preparation from individual hematopoietic colonies cultured in methylcellulose

T Amenomori, M Tomonaga, T Matsuo, Y Yoshida, K Kuriyama, N Sadamori and M Ichimaru

We report a micromethod for chromosome preparation from individual hematopoietic colonies cultured in methylcellulose. The entire process was carried out on poly-Lysine (PL)-coated slides. Individual colonies were transferred into 10 microliter of 0.075 M KCl and placed on PL- coated slides. After hypotonic treatment of the colony cells and their attachment to the slides, the cells were fixed by a three-step procedure as follows: addition of a 30% fixative (3:1 methanol:acetic acid) diluted with the hypotonic solution, addition of 20% ethanol, and subsequent immersion of the slides in a 100% fixative. The slides were flame dried and Giemsa stained. Q- and G-banding techniques also were used. These procedures provided analyzable chromosome preparations, even from colonies containing fewer than 50 cells.




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