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*10x Amino acid mix
*10x Amino acid mix
** See [[McClean:KS_Amino_Acid_Supplement]]
** See [[McClean:KS_Amino_Acid_Supplement ]]
Revision as of 17:21, 2 November 2011
Synthetic complete dropout media (SC-N with the nutrient N dropped out) is the media we use in the lab for selecting strains that have repaired an auxotrophy (e.g. selection on SC-URA plates after transforming with a URA+ plasmid or DNA fragment) or for persistent selection for the ability to grow without nutrient N (e.g. for maintaining a URA+ plasmid, the strain is always grown in SC-URA media).
Synthetic complete is the name given to the media with nothing dropped out (i.e. with all amino acids and other nutrients added the media). You can use synthetic complete for microscopy (it is much better for this purpose than YPD) but low fluorescence media (LFM) is preferable.
- Yeast Nitrogen Base without Amino Acids (Difco Cat #291940)
- Bacto-agar (Becton-Dickinson #214030)
- 10x Glucose Solution (20% w/v; Sigma D9434)
- 10x Amino acid mix (with appropriate nutrient dropped out)
- Distilled water
- 10x Glucose Solution (20% w/v)
For 1 liter of 20% glucose:
- Add 200g Dextrose (Sigma D9434) to 400ml of deionized water with with constant stirring. Add the water to the beaker first!
- Heat may help the sugar to dissolve
- Once the sugar has dissolved, bring the entire solution up to 1 liter. Divide into smaller bottles (usually you will use 50mls of 20% glucose to make 1L of media, so we usually aliquot about 100mls into small bottles). Autoclave for 30 minutes on the liquid cycle.
- 10x Amino acid mix
For 1 liter of media mix together:
- 6.7g yeast nitrogen base
- 20g bacto-agar
- deionized water to 900ml
Autoclave for 30 minutes on the liquid cycle. Once cooled (but see Notes) to about 55°C add 50ml of the 10x glucose solution and 50ml of the 10x amino acid mix. Pour plates.
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
- You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
- Anecdotal observations that might be of use to others can also be posted here.
Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
Burke, D. Dawson, D. & Sterns, T. 2000 Methods in Yeast Genetics: A Cold Spring Harbor Laboratory Course Manual Cold Spring Harbor Laboratory Press
- Megan N McClean 14:01, 20 July 2011 (EDT)
or instead, discuss this protocol.