====== Sporulation of Diploids ======

 --- //[[rylajosephine@gmail.com|Ryla Cantergiani]] 2020/03/30 17:07//

===== Overview =====
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Sporulation is how you generate haploid yeast strains from diploid yeast strains. An example of this would be to generate haploid deletion strains from a diploid strain from the het dip (heterozygous diploid) deletion collection.

If this doesn't work, try **GNA sporulation**, which takes longer but is reported to be more reliable.


===== Materials=====
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You'll need: 
  * Culture Tubes
  * YEP + 2% glucose
  * pipette tips
  * diploid strains of interest
  * 30°C shaker


=====Procedure=====
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  - Set up and label culture tubes. Label each tube with a piece of tape and tough spot.
  - Fill each culture tube with 3 mL of YEP + 2% glucose.
  - Inoculate each tube using a pipette tip (P10 or P20 work well) from a single colony of your diploid strain in question. Circle, initial, and date the colony used. Shake at 30°C overnight.
  - The next day, add 3ml of **Sporulation Media (SPM)** to a new culture tube for each strain (be sure to label your new tubes).
  - Using 1000μL tips and a P1000 pipettor, for each strain, aliquot 300μL of your diploid culture into the new tube containing 3mL of Sporulation Media. Shake at 30°C for 5-7 days before going to [[tetrad_digestion_and_dissection|Tetrad Dissection]]


=====Notes=====
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=====Part Numbers=====
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