Difference between revisions of "Experiment to determine whether yeast is contaminated and to salvage strain using 5FOA, 23Oct2014"

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(Created page with "'''October 23, 2014: Experiment to determine whether yeast is contaminated and to salvage URA- strain from possibly mixed sample''' *'''Location:''' BioCurious *'''Participan...")
 
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*'''Location:''' BioCurious
*'''Location:''' BioCurious


*'''Participants:''' Johan, Lafia
*'''Participants:''' Lafia, Johan


*'''Notes:''' Johan, Lafia
*'''Notes:''' Lafia, Johan


*'''Aims:''' Our yeast strains may be contaminated, so Rachel designed an experiment that would help us determine what strain we have and whether they are mixed. If indeed we have mixed or URA- strains, this experiment would allow us to pick out the URA- colonies so that we may use it. It would also give us confidence that we have a viable selection media to use when we do yeast transformations.
*'''Aims:''' Our yeast strains may be contaminated, so Rachel designed an experiment that would help us determine what strain we have and whether they are mixed. If indeed we have mixed or URA- strains, this experiment would allow us to pick out the URA- colonies so that we may use it. It would also give us confidence that we have a viable selection media to use when we do yeast transformations.
*'''Materials'''
** 12 plates from a new sleeve acquired at Biocurious
**  8 plates (4 each of YPD and YPD w/URA) pre-made by Teknova from a new sleeve from the batch ordered by Craig
** 250 ml YPD Agar + 5FOA, Prepared using Teknova YPD Broth from batch ordered by Craig. Agar was also from Teknova, ordered by Johan. 2.5 ml of 100x liquid 5-FOA was added after the media cooled below 45 degrees.
** 250 ml CM minus URA powder media ordered by Patrik (5FOA added after some of the media was used to make plates)
*'''Methods'''
***250 ml of Teknova broth and 5g of Agar was autoclaved in a glass bottle. Upon cooling below 45 degrees 5-FOA was added, as measured with a Extech IR thermometer.
***250 ml of CM minus URACIL liquid media was prepared using 8g of powder and 4g Agar.

Revision as of 17:52, 25 October 2014

October 23, 2014: Experiment to determine whether yeast is contaminated and to salvage URA- strain from possibly mixed sample

  • Location: BioCurious
  • Participants: Lafia, Johan
  • Notes: Lafia, Johan
  • Aims: Our yeast strains may be contaminated, so Rachel designed an experiment that would help us determine what strain we have and whether they are mixed. If indeed we have mixed or URA- strains, this experiment would allow us to pick out the URA- colonies so that we may use it. It would also give us confidence that we have a viable selection media to use when we do yeast transformations.
  • Materials
    • 12 plates from a new sleeve acquired at Biocurious
    • 8 plates (4 each of YPD and YPD w/URA) pre-made by Teknova from a new sleeve from the batch ordered by Craig
    • 250 ml YPD Agar + 5FOA, Prepared using Teknova YPD Broth from batch ordered by Craig. Agar was also from Teknova, ordered by Johan. 2.5 ml of 100x liquid 5-FOA was added after the media cooled below 45 degrees.
    • 250 ml CM minus URA powder media ordered by Patrik (5FOA added after some of the media was used to make plates)
  • Methods
      • 250 ml of Teknova broth and 5g of Agar was autoclaved in a glass bottle. Upon cooling below 45 degrees 5-FOA was added, as measured with a Extech IR thermometer.
      • 250 ml of CM minus URACIL liquid media was prepared using 8g of powder and 4g Agar.