Difference between revisions of "UW-Stout/Co Culture SP21"

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(Created page with "== This page is under construction == ==Materials== * double strength [https://sunrisescience.com/shop/growth-media/media-for-yeast/sd-complete-ynbnitrogen-glucose-csm-supple...")
 
(Protocol)
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==Protocol==
 
==Protocol==
 
# Create concentration of 100 cells per micro liter for both the knockout and control fluorescent strain with the Hemocytometer within 1 mL of double strength synthetic media.
 
# Create concentration of 100 cells per micro liter for both the knockout and control fluorescent strain with the Hemocytometer within 1 mL of double strength synthetic media.
# Place contents into a 2 mL culture within a deep well microassay plate
+
# Place contents into a 2 mL culture within a deep well microassay plate.
 
# Repeat steps 1 and 2 for each additional yeast strain, assigning each a unique well within the microassay plate.
 
# Repeat steps 1 and 2 for each additional yeast strain, assigning each a unique well within the microassay plate.
 
# Incubate the plate within the incubator shaker for 48 hours at 30 degrees Celsius, shaking at 600 rpm.
 
# Incubate the plate within the incubator shaker for 48 hours at 30 degrees Celsius, shaking at 600 rpm.
 
# From each well, take a 5 micro liter sample and mix it with 500 micro liters of phosphate buffered saline to measure on the flowcytometer.
 
# From each well, take a 5 micro liter sample and mix it with 500 micro liters of phosphate buffered saline to measure on the flowcytometer.

Revision as of 11:20, 6 April 2021

This page is under construction

Materials

Equipment

  • Shaking incubator set to 600 rpm and 30°C
  • flow cytometer
  • type 1000, 200, and 20 micro pipettors
  • Hemocytometer

Protocol

  1. Create concentration of 100 cells per micro liter for both the knockout and control fluorescent strain with the Hemocytometer within 1 mL of double strength synthetic media.
  2. Place contents into a 2 mL culture within a deep well microassay plate.
  3. Repeat steps 1 and 2 for each additional yeast strain, assigning each a unique well within the microassay plate.
  4. Incubate the plate within the incubator shaker for 48 hours at 30 degrees Celsius, shaking at 600 rpm.
  5. From each well, take a 5 micro liter sample and mix it with 500 micro liters of phosphate buffered saline to measure on the flowcytometer.