McClean: Intro to yeast: Difference between revisions
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(New page: <!-- COPY EVERYTHING BELOW HERE TO START YOUR OWN PROTOCOL! --> ==Overview== This introductory exercise is designed to get you started working with yeast. You will also take some data t...) |
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*Return the construct to the -80°C freezer as quickly as possible. '''DO NOT LET THE GLYCEROL STOCK THAW! REPEATED FREEZE THAW CYCLES WILL KILL THE STOCK!''' If you need to streak out multiple constructs take out only 1-2 from the freezer at a time or store them on dry ice while streaking. | *Return the construct to the -80°C freezer as quickly as possible. '''DO NOT LET THE GLYCEROL STOCK THAW! REPEATED FREEZE THAW CYCLES WILL KILL THE STOCK!''' If you need to streak out multiple constructs take out only 1-2 from the freezer at a time or store them on dry ice while streaking. | ||
*Use toothpicks to finish streaking the clone all over the plate so that you isolate single colonies. | *Use toothpicks to finish streaking the clone all over the plate so that you isolate single colonies. | ||
[[Image: | [[Image:Streak.For_.Single.Colony1.jpg ]] | ||
==Stock Solutions== | ==Stock Solutions== | ||
Revision as of 07:45, 27 April 2015
Overview
This introductory exercise is designed to get you started working with yeast. You will also take some data that will be useful to you anytime you are working with yeast in the lab.
Streaking Yeast out from Glycerol Stocks
- Wipe down your bench area with 70% ethanol to give yourself a clean workspace.
- Based on the yMM number, determine the location of the construct you want to streak out. It will be located in the -80°C freezer. Know where you are looking before you open the freezer door, sitting around with the door open is a no-no.
- Take the tube back to your bench. Ideally, store it on dry ice while streaking or be really quick.
- Take a sterile applicator stick. Scrape off a portion of the top of the frozen glycerol stock and streak it onto the edge of your plate.
- Return the construct to the -80°C freezer as quickly as possible. DO NOT LET THE GLYCEROL STOCK THAW! REPEATED FREEZE THAW CYCLES WILL KILL THE STOCK! If you need to streak out multiple constructs take out only 1-2 from the freezer at a time or store them on dry ice while streaking.
- Use toothpicks to finish streaking the clone all over the plate so that you isolate single colonies.
Stock Solutions
Stock Solution 1
- This is a very simple solution, so we only need a one line description of how to make it.
Stock Solution 2
This is a more involved solution, so we will describe how to make it in several steps:
- Step 1
- Step 2
- Step 3
Protocol
- Step 1
- Step 2
- Step 3
Notes
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.
References
Gietz, R.D. and R.A. Woods. (2002) TRANSFORMATION OF YEAST BY THE Liac/SS CARRIER DNA/PEG METHOD. Methods in Enzymology 350: 87-96.
Contact
- Megan N McClean 14:01, 20 July 2011 (EDT)
or instead, discuss this protocol.
