Manually setting 96 wells plates with lower volume samples!

From: xaravich ivan
Date: 18 November 2011 14:34


Hi,
I apologize in advance as it is not a ccp4 related question, but over the years, CCP4bb is synonymous with protein crystallographers virtual university, at least for me.

Ok, now I do not have an easy access to crystallization robot, so I was hoping if someone here have ever used the 96 well plates for manually setting drops with much lower solution/sample volumes (0.1-0.2micro litres).

I heard about a clicker syringe that can be used to manually add lesser volumes but I am not sure how to go about it.
Please let me know if you are aware of or  are routinely setting 96 well trays with much lower volumes of crystallization solutions as well as samples, manually.

thanks in advance,
ivan

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From: Shya Biswas

Hi,
I am routinely using the gryphon robot from Art Robbins.This instrument can dispense 0.02microlitre at minimum. Your drops can dry out if you use such low volumes you have to be really fast. You can set up 0.2 to 0.1 micro litre drops using this for 96 well plates however the instrument needs to be calibrated well sometimes with low volume drop the robot dispenses the protein at one corner of the well and the precipitant at another end so eventually the outcome is the mixing of the precipitant with your protein is not happening. However with 0.2micro litre drop or higher this is less of a problem.
HTH,
shya

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From: Shya Biswas


sorry totally misunderstood your question, however if you can ship your protein i can always try to setup a tray for you.
Shya

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From: Ed Pozharski


The best we could do was to use the multichannel pipettor that goes down
to as little as 0.5ul to add reservoir solution to the drop and a
repeater (that's what you probably mean by "clicker syringe").  The
smallest volume you can dispense with a repeater is 1ul, but it has the
advantage of speed (no need to go back into the protein tube) and
positive displacement mechanism (no worries about viscosity/surface
tension).

So one cannot get much advantage regarding protein amount, it still
takes ~100ul to set up a 96-well tray (which robotic systems can bring
down by an order of magnitude).  But you do save a lot of money on the
screens and the methodology is much faster than traditional dispensing
(with screening solutions in deep well blocks the whole dance takes 3-4
minutes).

Cheers,

Ed.

--
"Hurry up before we all come back to our senses!"
                          Julian, King of Lemurs

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From: Martin Hällberg


Hi,
With the electronic Biohit 8 channel pipette you can set up 3x96 drops in 5 minutes. Lowest volume is 0.2 ul. We have used this system for a couple of years with RNase free filter tips for RNA-protein complexes. With the recent arrival of a shared Mosquito we have used the Biohit less but it is still convenient to have the setup. You need good eyesight and a steady hand, though.

Cheers,

Martin

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From: Ho Leung Ng k


Hello Ivan,

    You can dispense 0.1-0.2 uL drops with the PB-100 repeating
dispenser from Hamilton and a 10 uL glass syringe. But it's rather
clumsy and hard on the hands. I don't recommend using it with 96 well
plates. It's too slow to do the whole plate at one time without your
drops drying up.


Ho

Ho Leung Ng
University of Hawaii at Manoa
Assistant Professor, Department of Chemistry

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From: David Salom


it can be done but, if you don't want your drops to dry, better do it in a cold room. It's painful though.

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From: Jacqueline Vitali


I have the same problem as I am not using a robot.  

For this reason I use the 48 well hanging drops from Hampton and seal them one at a time.  It is very time consuming but uses little reservoir solution and your drops do not dry out..

Jackie Vitali