Team:UNICAMP-Brazil/Notebooks/October 20

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(Yeast experiments: Lysozyme)
(Yeast experiments: Lysozyme)
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*<p style=”text-align:justify;”>Today we began the tests with the yeasts transformed with the following constructions: YEP+pDLD-Lysozyme and YEP+Adh1-Lysozyme.</p>
*<p style=”text-align:justify;”>Today we began the tests with the yeasts transformed with the following constructions: YEP+pDLD-Lysozyme and YEP+Adh1-Lysozyme.</p>
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*<p style=”text-align:justify;”>We inoculated the yeast pre inocula in 3 types of media: YNB Glucose Ura-, YNB Ethanol Ura- and YNB Glucose Ura+. The inocula grew for 4 hours in 30ºC and 150rpm before the induction with lactic acid. Four concentrations of lactic acid were used in the induction.</p>
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* The pre inocula made yesterday grew. We inoculated it in 50mL of YNB Glucose Ura- or YNB Ethanol Ura-. The wild type yeast is Ura- so we grew it on YNB Glucose Ura+. The inocula grew for 4 hours in 30ºC and 150rpm until the OD 1,3 before the induction with lactic acid.  
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* We performed the induction 5 hours long. We mesured the OD every one hour to construct a growth curve. At the end of the 5 hours we pelleted the culture, colected the supernatant and freezed the pellet to analyze in SDS-PAGE tomorrow.
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* We also dripped drops of the supernatant in filter paper discs and placed them in a ''Lactococcus lactis'' plate. We hope to see inhibition zones surrounding some of the filters.
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* The Yeasts inoculated in YNB Ethanol media didn't grow enough to begin the induction until the end of the day.  

Revision as of 22:23, 21 October 2009

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Yeast experiments: Lysozyme

  • Today we began the tests with the yeasts transformed with the following constructions: YEP+pDLD-Lysozyme and YEP+Adh1-Lysozyme.

  • The pre inocula made yesterday grew. We inoculated it in 50mL of YNB Glucose Ura- or YNB Ethanol Ura-. The wild type yeast is Ura- so we grew it on YNB Glucose Ura+. The inocula grew for 4 hours in 30ºC and 150rpm until the OD 1,3 before the induction with lactic acid.
  • We performed the induction 5 hours long. We mesured the OD every one hour to construct a growth curve. At the end of the 5 hours we pelleted the culture, colected the supernatant and freezed the pellet to analyze in SDS-PAGE tomorrow.
  • We also dripped drops of the supernatant in filter paper discs and placed them in a Lactococcus lactis plate. We hope to see inhibition zones surrounding some of the filters.
  • The Yeasts inoculated in YNB Ethanol media didn't grow enough to begin the induction until the end of the day.


Raíssa and Taís