流式检测细胞周期时PI与PNA酶究竟要加多少的量

Question

如题所述

Answer 1

贴一个通用的protocol：

Harvest the cells in the appropriate manner and wash in PBS.

Fix in cold 70% ethanol. Add drop wise to the pellet while vortexing. This should ensure fixation of all cells and minimize clumping.

Fix for 30 min at 4°C.

Wash 2 X in PBS. Spin at 850 g in a centrifuge and be careful to avoid cell loss when discarding the supernatant especially after spinning out of ethanol.

Treat the cells with ribonuclease. Add 50 µl of a 100 µg/ml sock of RNase. This will ensure only DNA, not RNA, is stained.

Add 200 µl PI (from 50 µg/ml stock solution).