McClean:YPD2: Difference between revisions

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(New page: ==Overview== Yeast nitrogen base without amino acids with agar for plates. This protocol leaves 100ml (per L) of room for additives (glucose, amino acids, etc). Media is labelled as YNB1...)
 
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==Overview==
==Overview==
Yeast nitrogen base without amino acids with agar for plates.  This protocol leaves 100ml (per L) of room for additives (glucose, amino acids, etc).  Media is labelled as YNB1 on the media shelf.
YPD agar for plates.


==Reagents==
==Reagents==
* Yeast Nitrogen Base (without amino acids)
* Bacto yeast extract
* Bacto Agar
* Bacto Peptone
* MilliQ H<sub>2</sub>O
* MilliQ H<sub>2</sub>O
* Dextrose


==Protocol==
==Protocol==
*Container 1:
*Begin with approximately 700mls of MiliQ water
** Begin with approx. 700ml MilliQ H<sub>2</sub>O in 2L bottle
*Weigh out and add ingredients with constant stirring:
** Add with constant stirring: 20g of Bacto Agar
**Bacto yeast extract 10g
** Bring up to 800ml with MilliQ H<sub>2</sub>O
**Bacto peptone 20g
** Autoclave
*Weigh out and add with stirring:
** Allow to cool (50°F)
*Bacto agar 20g
 
*Bring up to 900mls with MiliQ water
*Container 2:
*Separate YP into two 1L bottles of 450mls each
** Begin with approx. 80ml MilliQ H<sub>2</sub>O
*Weight out 20g of dextrose and add to 60mls of MiliQ water and bring volume up to 100ml total
** Add with constant stirring 6.7g of Yeast Nitrogen base (without amino acids)
*Autoclave separately
** Bring up to 100ml with MilliQ H<sub>2</sub>O
*Allow to coole in 55°C water bath
** Autoclave
*Combine each 450mls YP with 50mls of glucose solution
** Allow to cool (50°F)
 
Combine contents of 2 containers and mix well


==Notes==
==Notes==

Revision as of 10:13, 2 July 2015

Overview

YPD agar for plates.

Reagents

  • Bacto yeast extract
  • Bacto Peptone
  • MilliQ H2O
  • Dextrose

Protocol

  • Begin with approximately 700mls of MiliQ water
  • Weigh out and add ingredients with constant stirring:
    • Bacto yeast extract 10g
    • Bacto peptone 20g
  • Weigh out and add with stirring:
  • Bacto agar 20g
  • Bring up to 900mls with MiliQ water
  • Separate YP into two 1L bottles of 450mls each
  • Weight out 20g of dextrose and add to 60mls of MiliQ water and bring volume up to 100ml total
  • Autoclave separately
  • Allow to coole in 55°C water bath
  • Combine each 450mls YP with 50mls of glucose solution

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

  1. List troubleshooting tips here.
  2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
  3. Anecdotal observations that might be of use to others can also be posted here.

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References

Contact

  • Who has experience with this protocol?

or instead, discuss this protocol.


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