Protocol 31 - Primary culture of embryonic cells (Kevin Strange and Rebecca M)


I’ doing a modified version of killing assay ( ) and need av way to separate eggs from debris. This model: Methods in cell biology - protocol 31 describes a way to separate egg and debris, but the method does not describe how many plates should be used per 15 ml tube in order to separate eggs and debris completely.

Since I’m using a killing assay its crucial for me to have a large output of synchronization. Have anyone done this separation technique before and how many synchronized plates per 15 ml tube did you use?

I’m grateful for every reply!

Kjetil F.C
Oslo University Hospital


not sure why you need protocol 31 as that is really for isolating embryonic cells rather than for synchronising worms.

Here is a link to the Petrascheck Lab paper (Greg Solis) where they describe (among other things) how they synchronise their worms, how many plates, tubes, worms etc…it’s also describes a lifespan assay in a 96 well plate, so the worm numbers, volumes analysis might also help you;

It’s very detailed and there’s a free to view bonus video!




Thanks for your reply steveh. I’ve looked trough the protocol you linked to and can’t find that they describe a method for separating debris and egg - witch is my aim. In the protocol they wash the worms 3 times and centrifuge between each time after bleach is added. From my experience this is not enough to remove all the debris as this also will pellet.

Although protocol 31 is for embryonotic cells I’m wondering if anyone had done this as part of synchronization with a high output result, if yes - how many plates did you use per 15 ml tube?

I’m thinking that when centrifuging 15 ml tubes with debris and eggs - if you use too many plates per 15 ml tube the 30% sucrose solution will not be able to separate and the eggs will fall down and pellet.

  • Kjetil


the key point post-bleach is that one should resuspend the egg pellet for each of the wash cycles. If you don’t, you get lots of debris.

I have just done a egg prep using this method and used the eggs to photograph promoter::gfp reporter expression in different stage embryos…I had no debris.

As for the numbers of plates, I hope someone can help you further…