seperations

[musiclink213]

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Anyone know how to seperate 2 layers that form after centrifugation? everytime i try, i always end up leaving a lot of it in the tube, or getting a lot of the top layer into the solution, and then my lab results get all screwed up. anyone have any tips for someone like me who cannot seperate? i've tried everything! even practicing at home with oil and water and trying to seperate that, but nothing helps!

 

[Gleevec]

Originally posted by musiclink213
Anyone know how to seperate 2 layers that form after centrifugation? everytime i try, i always end up leaving a lot of it in the tube, or getting a lot of the top layer into the solution, and then my lab results get all screwed up. anyone have any tips for someone like me who cannot seperate? i've tried everything! even practicing at home with oil and water and trying to seperate that, but nothing helps!

Can you use phase-lock gel caps? They have a gel interface that causes your micropipette tip to get clogged if you go too deep, and the phases seperate pretty readily.

If you cant do that, just use a bigger micropippetter initially, then switch to a smaller one. Better to use up a couple more tips than to pick up some junk.

 

[metabolite]

I have some lab experience and have pipetted a few stuff in the past 6 years or so... depending on what you are trying to separate, but since you are noticing that you cannot separate well I am assuming that samples are actually visible (i.e. blood perfusate, homogenate extract, something in that area) and what you are separating happens to be both at liquid phase (oil/water example)

Most of the times if things don't separate well that has to do with g-value at which you are centrifuging... may be the speed is not fast enough to effectively form distinct layers.

If that's not the case, use smallest tip you can use and pipette SLOWLY.

If you are separating two solvents, you might have to use some anhydrous material to dry out liquid phase - which I'm not an expert.

If you are afraid that what you have pipetted may be contaminated with substances/solvent that you didn't want, can you run it through SPE? You might be able to purify the sample that way.

I hope it helps.