PCB 3063 Lecture Notes - Lecture 1: Photographic Emulsion, Thymidine, Autoradiograph
De novo
- starts from the beginning = doesn't require a primer
- ex: RNA synthesis can be "de novo"; DNA synthesis can't be
Taylor, Woods, Hughes experiment
- pulse chase style on beans
- used ³H thymidine
Procedure:
- grow beans in radioactive ³H thymidine
- switch to cold (non radioactive) thymidine
- stop cells at metaphase
- squash cells and dip them in photographic emulsion
Findings:
- inactivated X replicated last
- euchromatin replicates early
- heterochromatin replicates late
- homologous recombination
Autoradiography
image of radioactive particles as they degrade
Pulse-Chase experiment
pulse = cells are exposed to a labeled (ex: radioactive) compound
chase = cells are in normal version of that compound
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