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Re: [ccp4bb] About seeding in microbatch under oil crystallization
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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999
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Subject: Re: Native Gel Theory and Practice
From: Jürgen_Bosch jubosch {- at -} JHSPH {- dot -} EDU
Date: 2010-05-19		Next message:
Subject: Re: About seeding in microbatch under oil crystallization
From: Ravindra Makde ravimakde {- at -} YAHOO {- dot -} COM
Date: 2010-05-19


Subject: Re: About seeding in microbatch under oil crystallization
From: wu donghui wdh0972 {- at -} GMAIL {- dot -} COM
Date: 2010-05-19

Dear Patrick,

I have Oryx robot in our lab. I got my initial crystallization hit from
sitting drop vapor diffusion method. This hit can be reproduced by sitting
drop through spontaneous nucleation with many tiny crystals after around 1
week but seeding into sitting drop failed and I found once I opened slide
and let the drop exposed to air even for a few second, it will turn turbid
instantly and I think this might be the reason why seeding in sitting drop
is not workable as most protein precipitated and protein concentration
dropped very low and seeds implanted inside might be dissolved quickly even
after certain periods of equilibration before seeding. That is the reason I
want to seperate my drop from air by adding oil on it and then try seeding
under oil. Also microbatch under oil might prevent skin formation and
maintain protein concentraiton at relatively high level to support seed
crystal growth.

Best regards,

Donghui

On Wed, May 19, 2010 at 7:37 PM, Patrick Shaw Stewart > wrote:

> Dear Donghui
>
>
>
> I can confirm that microseeding with microbatch-under-oil works fine.
>
>
>
> It’s one of our standard set of scripts. Our Oryx robots use contact
> dispensing (the tip touches the plate) so the seeds can be added very
> reliably as a suspension.
>
>
>
> Several users have reported that the method works, although I don’t know of
> anyone who has made a statistical comparison between Sitting Drop and
> Microbatch. I would expect them to be about the same.
>
>
>
> Do you have an Oryx or some other robot? And was there any particular
> reason why you want to use microbatch?
>
>
>
> Best wishes
>
>
>
> Patrick
>
>
>
>
>
>
>
>
>
>
>
> --
>
> For information and discussion about protein crystallization and
> automation, please join
>
> our bulletin board at http://groups-beta.google.com/group/oryx_group?hl=en
>
>
>
> patrick@douglas.co.uk Douglas Instruments Ltd.
>
> DouglasHouse, EastGarston, Hungerford, Berkshire, RG177HD, UK
>
> Directors: Peter Baldock, Patrick Shaw Stewart
>
> http://www.douglas.co.uk/
>
> Tel: 44 (0) 148-864-9090 US toll-free 1-877-225-2034
>
> Regd. England 2177994, VAT Reg. GB 480 7371 36
>
>
>
> *From:* CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] *On Behalf Of *wu
> donghui
> *Sent:* 19 May 2010 10:29
>
> *To:* CCP4BB@JISCMAIL.AC.UK
> *Subject:* [ccp4bb] About seeding in microbatch under oil crystallization
>
>
>
> Hi ccp4bbers,
>
>
>
> I want to know if anyone has any experience about seeding (streak seeding
> or microseeding) in microbatch under oil crystallization. I wonder if the
> oil might block or wipe away the seeds if cat whisker is used for streak
> seeding. Thank you for your input ahead.
>
>
>
> Best regards,
>
>
>
> Donghui
>
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