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Attractant Media - labwiki.html
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<h1 id="firstHeading" class="firstHeading" lang="en"><span dir="auto">Attractant Media</span></h1>
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<div id="siteSub">From labwiki</div>
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<div id="mw-content-text" dir="ltr" class="mw-content-ltr" lang="en"><p>Thoughts on making heterogenous media for experiments where we want to localize worms to a specific region of the plate.
</p><p><br>
</p>
<h2><span class="mw-headline" id="Choice_of_Attractant">Choice of Attractant</span><span class="mw-editsection"><span class="mw-editsection-bracket">[</span><a href="http://ecotheory.uwaterloo.ca/labwiki/index.php?title=Attractant_Media&action=edit&section=1" title="Edit section: Choice of Attractant">edit</a><span class="mw-editsection-bracket">]</span></span></h2>
<p>Some work by an iGEM team (<a rel="nofollow" class="external text" href="http://2013.igem.org/Team:Valencia_Biocampus">iGEM team</a> <a rel="nofollow" class="external text" href="http://www.wormbook.org/chapters/www_behavior/behavior.html#sec5">WormBook Attractants</a> suggests that MgSO4 is a reasonable attractant for <i>C. elegans</i>,
though preliminary tests in this lab were not very promising. An ideal
attractant should be soluble in agar media so that it can be
incorporated into defined regions of the plate.
</p><p><br>
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<h2><span class="mw-headline" id="Media_Preparation">Media Preparation</span><span class="mw-editsection"><span class="mw-editsection-bracket">[</span><a href="http://ecotheory.uwaterloo.ca/labwiki/index.php?title=Attractant_Media&action=edit&section=2" title="Edit section: Media Preparation">edit</a><span class="mw-editsection-bracket">]</span></span></h2>
<p>Media Recipe (per Litre)
</p><p>NaCl - 3g
</p><p>Agar - 17g
</p><p>Peptone - 2.5g
</p><p>1M CaCl2 - 1mL
</p><p>5mg/mL Cholesterol - 1mL
</p><p>1M MgSO4 - 1mL
</p><p>1M KPO4 - 25mL
</p><p><br>
To make a heterogenous plate, prepare two sets of media, one following
the above recipe, and one more following the above recipe + your
attractant of choice. Once your media are both ready, prepare to pour
the plates. You'll need the following:
</p><p>As many plates as you'd like to poor.
</p><p>A metal test tube cap, must be metal and must have a relatively sharp edge.
</p><p>Tweezers.
</p><p>Ethanol flame/burner.
</p><p>Ethanol jar for flame sterilization.
</p><p>P1000 pipette & tips.
</p><p><br>
Obvious instructions regarding aseptic technique will be left out of the
following description, be sterile! First, pour the normal agar in to a
plate, fill the plate as much as you'd like. Let the normal agar set
fully, shouldn't take more than 10-15 minutes. Once the normal agar is
set, ethanol flame sterilize the metal test tube cap, holding it
securely with the tweezers. Once the cap has cooled enough to hold with
your gloved hand, grab it by the top (closed end, keep the metal edge
sterile). Press firmly into the set normal agar with the edge of the
metal cap, the agar should cut, not crack, twist a little if you need
to. Once the metal cap is all the way through the agar, to the bottom of
the plate, angle it slightly to pop the disk of agar out. It may be
necessary to use the tweezers to remove the agar disk. With the disk
removed, use the P1000 to fill the empty region with your special
attractant media, making sure to fill the hole as completely as
possible. Any cracks or edges will give the worms added cause for
burrowing. Let the added agar set.
</p>
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