Re: [ccp4bb] crystallisation

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CCP4bb <-- 2008 <-- June 2008 <-- 04 June 2008

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Subject: Re: Crystals grown directly on loop?
From: "Mathews, Irimpan" iimathew {- at -} SLAC {- dot -} STANFORD {- dot -} EDU
Date: 2008-06-04

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From: Paul Adams PDAdams {- at -} LBL {- dot -} GOV
Date: 2008-06-04

Subject: Re: crystallisation
From: sajid akthar b_sajid_ali {- at -} YAHOO {- dot -} CO {- dot -} IN
Date: 2008-06-04

Dear All

Thank you for your suggestions. I got many suggestions
from you. Due to my time constraint, I'm not writing
summary, but some replies for your suggestions to my
previous query.

I have not mentioned fully what I tried to do better
diffraction. I'm listing them here which were
mentioned by some of you.

I have tried the following cryo protectans.

1) Glycerol 5-30%. Best is 10%. And I have tried
serially transferring crystal from lower amount of
glycerol to higer Glycerol. This did not worked well.

2)Sucrose as cryo protectant. The same experiment was
performed as in glycerol and not better than
10%glycerol alone.

3)Glutaraldehyde as cryo protectant. And all the
experiments mentioned in the reference of Acta D61,
1173-1180. NO good result. Annealing experiment with
my crystal did not help.

Yet I have not tried to thaw crystal (from N2
stream)back into cryo cnodition and again check for
diffraction. Also I have not tried collecting at room
temperature.

4)ParatoneN oil alone and paratone with glycrol,
Paratone with sucrose were tried as cryo condition.
Mosaicity was too much.

5)Immersion oil TypeA combination with 10% glycrol
gives some lower mosaicity. Still not great from 10%
glycerol

6)Hampton screen additives did not give any crystals
apart from the original condition

7)Silver Bullet screen too failed to produce crystal
at any well.

8)No crystal growth at 5 degrees.

9)I'm not sure about Phoebe Rice idea. And I did not
checked that too.

10) DTT is fresh in the sample and it is 1mM.

Thank You

Sajid

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CCP4bb <-- 2008 <-- June 2008 <-- 04 June 2008

Previous message:
Subject: Re: Crystals grown directly on loop?
From: "Mathews, Irimpan" iimathew {- at -} SLAC {- dot -} STANFORD {- dot -} EDU
Date: 2008-06-04

Next message:
Subject: System Administrator Position in Berkeley California
From: Paul Adams PDAdams {- at -} LBL {- dot -} GOV
Date: 2008-06-04