(b) Experimental Approach for Testing Rvl2 in Yeast (30%)
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14 Terms
What is the goal of the experimental approach?
To determine if Rvl2 prolongs lifespan and to identify its mechanism of action. (b)
What organism is used for testing?
Yeast.
Why is Chronological Life Span (CLS) the better model?
Because Rvl2 is intended for human use, and CLS mimics post-mitotic aging (like human neurons). (deprez) (cruz-bonilla)
How is yeast grown in the CLS assay?
Grow yeast to stationary phase in 96-well plates. (deprez) (cruz-bonilla)
How is viability measured periodically in CLS?
Via outgrowth (OD600) monitoring or Phloxin B staining. (deprez) (cruz-bonilla)
What does Phloxin B stain?
It only enters and stains dead cells. (deprez) (cruz-bonilla)
What method is used for Replicative Life Span (RLS) assay?
Microdissection or automated micro-fluidic "micro-buckets". (b)
What does the RLS assay count?
The number of daughter cells produced by individual mother cells. (b)
What two conditions are compared in RLS?
Mother cells treated with Rvl2 versus a control. (b)
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Experimental Approach for Testing Rvl2 in Yeast (30%)
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Testing for Lifespan Extension
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Chronological Life Span (CLS) Assay
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Replicative Life Span (RLS) Assay