BIOB11H3 Lecture Notes - Lecture 15: G2 Phase, Heterokaryon, S Phase

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3 Mar 2017
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Mitosis: division of nuclear material // cytokinesis: division of cytoplasm. Consider an asynchronous cell culture (cells in different cell cycle, some in g1, some in s, some in m etc) Add 3h-thymidine, label 15 min, fix and conduct autoradiography / count cells which are radiolabelled. 9/30 cells are labelled 30% are in s phase 30% of the cell cycle is in s phase. *dna replication occurs in s phase hence 3h-thymidine will be incorporated into dna. Asynchronous cells: label with radioactive dntp for short time incorporation of label only occurs in s phase. Examine cells at later times for mitotic chromosomes (condensed) check are the mitotic chromosomes labelled. *the first mitotic chromosomes to appear will not be labelled it was far beyond s phase and no new dntp* will be incorporated. Add 3h-thymidine for 30 minutes to an asynchronous culture. Time between start of labeling period to appearance of the first labelled mitotic chromosome.

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