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I attempted my first yeast washing last night after bottling. I added 1 quart of boiled-then-cooled water to my carboy, swished the yeast cake into suspension, then filled 2 quart jars, each about 3/4th's full. After about 30 minutes there was maybe 1/2" of seperation. At this time, I really needed to get to bed so I let it sit overnight. In the morning (about 7 hours later) there was a lot more seperation, but the top layer was much clearer than any of the videos I've watched online.

My question is, is it possible too much of the yeast fell out of suspension? Also, I'm confused if there is anything else that needs to be in the jars besides water and yeast cake. Does it need nutrients?

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I don't think there is any danger in the timing, but if your sample contained a lot of trub, having it all settle out with the majority of the yeast defeats the purpose of the wash. If you can see two distinct layers formed in the bottom you likely have a lot of trub in there and I'd suggest agitating things again so you can pour off the yeast while it's still in suspension.

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Ok, so based off this comment and some other things I've read, I guess there would be 3 layers not 2 as I assumed. The top section that I was referring to (which is nearly clear) can be discarded. I think I have very little trub, I did see a small delineation but only about 5% was in the lowest section. Is that feasible? –  user2101 Aug 8 '12 at 20:52
    
If your trub layer is minimal, you're probably okay. Some people pitch right onto the previous cake so they aren't worried about it. The biggest thing is if the style you're brewing is "delicate" there is more of a chance of picking up coloring/flavoring from the trub. If you're pitching into a 10% imperial stout, or a double ipa then you're less likely to notice anything from the previous brew. –  DHough Aug 9 '12 at 14:41
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No, there is no danger in leaving it longer and getting more separation. If anything, it means you get even more yeast, although the amount is only a few percent.

If there is a lot of trub, then you may want to pour off the yeast into a different vessel to separate it from the trub. The trub falls quickest, so this will be at the bottom of your jar.

Store the jar in the fridge. No need for nutrients. You can pitch directly if used within 2 weeks. Longer than that and it's best to make a starter and pitch the actively fermenting yeast.

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