Lab Six

What is a spot assay?

A method usd to compare growth of wild type and gene fusion yeast strains.

What does a spot plate show on a rich media?

It allows comparison of growth rates in ideal conditions

What does a spot plate do on a specific media?

It shows the function of the gene under conditions like salt or an alternative carbon source.

In today’s lab what are we doing?

Preparing Serial Dilution and Plating onto Stress Media

What are the goals in today’s lab?

To observe changes in expression of your GOI under different environmental conditions, and understand GOI responds to environmental stress. 

Why is the stop codon removed?

To ensure GOI and GFP are translated into one proteins.

How are we testing differences between experimental and negative control groups?

Test with spot dilution plates

What kind of serial dilution is done in this lab?

1:10

What is Sterile Technique?

Wiping down the lab bench with 70% ethanol, working near a flame, and sterilizing reagents in auto clave and using sterile pipettes

In this lab what is osmotic stress?

1M NaCl concentration

In this lab what is Oxidative stress?

7 mM hydrogen peroxide

In this lab what is Metal toxicity?

15 mM CuIISO4 concentration

In this lab what is Ethanol treatment?

10% ethanol

In this lab what is Alternative carbon source?

: 3% glycerol in place of glucose in YPD

In this lab what isTemperature?

cold (expose to 20 °C) and heat (expose to 50 °C) shock for 30
min

In this lab what is DNA damage sensitivity?

UV-light. Immediately after spots have dried, expose
the spots to a UV light box (agar side down) for 1 min. Wrap in foil for incubation to prevent light-dependent DNA repair