# Title: 22082019 Mating using liquid drops of yEK7a and ylic133_5 💔

# Date

22082019

# Objective

Goal: To try to achieve a more efficient mating between ylic133 and yEK7a, because the previous attempt did not get good results after sporulation.

# Method

  • Grow an overnight culture of each of the strains in 10mL YPD
  • Next morning take a YPD agar plate and add 20ul of each dense liquid culture one on top of each other.
  • Microscopy time lapse imaging of the mixture of both strains 100ul of 150X yEK7a (highly dense) and ylic133_5 (no dense at all) no diluted , after being washed in non fluorescence media (CSM).
  • 23082019-Restreaking of the mated patch onto a YPD+G418+NAT+Hygro plate.
  • 26082019-Inoculation of 8 restreaked cells to sporulation next morning.

# Results

  • There are "weird structures" on the ylic133_5 well, flowers like structures. It could be some sort of contamination, but which type?
  • Jut one cell appear to show this dumbell shape (top left last frames) 😦
  • As a next step , I will image all my glycerol stocks from ylic133 and ylic132
  • ylic133
  • ylic132

# Conclusion

  • I dont see these weird structures in the images of my stock meaning that what I saw during the mating was maybe something acquired outside the glycerol stock.