McClean: Tetrad Dissection

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
Line 36: Line 36:
<!-- Please paste this section "as is" into your protocol, and add notes to it if you have some!-->
<!-- Please paste this section "as is" into your protocol, and add notes to it if you have some!-->
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
-
#List troubleshooting tips here. 
+
#'''*[[User:Megan N McClean|Megan N McClean]]''' When I can't age my dissection plates, I will stick them in the 30°C or 37°C warm room the morning of the day I dissect to dry them out a little bitIt is absolutely infuriating, if not impossible, to try dissection on plates that are wet.
-
#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 <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.
Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.

Revision as of 13:12, 30 September 2011


Contents

Overview

This protocol describes dissection of yeast tetrads. In our lab, we primarily use tetrad dissection for constructing strains for genetic and biochemical experiments.

An accomplished yeast biologist can dissect a plate of spores in 20 minutes; for a beginner, 2 hours is not unusual. Tetrad dissection is a learnable skill. Your initial attempts will likely be frustrating. If you persevere, you will be richly rewarded by your new ability to wield one of the most powerful tools in the yeast geneticist’s toolbox. When you first learn to do tetrad dissection, make sure to ask for help from someone in the lab who is experienced at doing it! It really helps to have someone with a practiced eye point out what a well-digested culture looks like, what a tetrad looks like under the dissecting scope, etc.

Materials

  • β-glucuronidase (Sigma G7770, stored in 4°C refrigerator)
  • Sporulated yeast culture
  • Sterile Water
  • "Dry" YPD dissection plates
    • Everyone has a particular way that they like their plates for tetrad dissection. You are basically aiming for dry and level. To make dissection plates, add 25 ml YPD with a plastic strippette to plates on a very level surface. Once solid, invert. Let dry at room temperature for ~3 days. Bag. These plates are best after aging for a while.


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:

  1. Step 1
  2. Step 2
  3. Step 3

Protocol

  1. Step 1
  2. Step 2
  3. Step 3


Notes

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

  1. *Megan N McClean When I can't age my dissection plates, I will stick them in the 30°C or 37°C warm room the morning of the day I dissect to dry them out a little bit. It is absolutely infuriating, if not impossible, to try dissection on plates that are wet.

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

or instead, discuss this protocol.

Personal tools