Hi everyone,
I have done chromosome staining in mice bone marrow with colchicine treatment and stained using giemsa 5 %. But I think it did not work. Here are my steps:
1. Mice was injected by colchicine intra peritoneal ( 0,1 %, 1ml / 100 g body wt) 2 hours before killing the mice
2. Mice was killed and bone marrow cell were flushed from the femora with PBS 1x then centrifuged at 1000 rpm for 5minutes and added 200 ul 0,56 % KCl into pellet for 30 minutes at 37 celcius.
3. Then I centrifuged using a same rpm and remove supernatant. Pellet was added with 200 ul carnoy fixative (methanol:glacial acetic acid, 3:1). Resuspended and centrifuged.
4. Removed supernatant and pellet was added with fresh carnoy fixative the resuspended.
5. I smeared on clean glass slides and left to dry. then stained with giemsa.
The result was not good at all. What was wrong?
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