Hi Paul,
sometimes the simplest methods are the best. This one is from field
nematologists and is called the "Baerman funnel".
Attach a rubber tube to the end of a funnel and clamp it closed with
a pinch-clamp or screw-down type "C" clamp. in the mouth of the
funnel, put a small screen, which acts as a support for a filter. But
instead of filter paper, use two KimWipes. Wet the KimWipes by
squirting water into the funnel, and remove the air bubbles that
invariably get caught in the tubing (e.g., by squeezing the tubing
several times or opening the clamp to let some water down into the
tube).
Put your worms + agar (or worms that have burrowed into agar, or
worms in dirt, or live worms in any matrix) onto the KimWipes. Add
water to just cover them. Over a period of a couple hours, the worms
will crawl out of the agar pieces and through the mesh of the
KimWipes. They will then sink to the bottom of the clamped tubing.
Collect the worms by opening the tubing slightly and letting a few
drops of water drip into a test tube. Your worms will have no agar
or dirt.
Another method is to float the worms on 30% sucrose and centrifuge.
Agar will generally pellet faster than the worms. But the worms are
all shriveled up.
Dave
>I am a researcher currently attempting to perform in situ
>hybridisation experiments on adult C. elegans. Unfortunately, the
>experiments are being hampered somewhat by the presence of agar (from
>the growth plates) in the reaction mixture.
>>Can anyone give me a RELIABLE protocol for removing agar fragments
>from the worms?
>>Thanks a lot,
> Paul McVeigh
> Parasitology Research Group
> Queens University Belfast
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