I've always found it is a much more effective and delightful use of my
time to mope sullenly in my office, communicate with my family members and
colleagues while restricting my working vocabulary to monosyllabic grunts,
or pensively pace the halls, inflicting my abject misery upon hapless
passers-by, but then I crystallize RNA.
On Sun, October 11, 2009 12:01 pm, [log in to unmask] wrote:
> depending on the importance of the project, I would be tempted to just
> wait for the crystals to grow, and in the meantime:
> - work on another project
> - start a new project
> - write a grant application, a paper, a review...
> or a combination of the above.
> Mark
>
> Quoting William Scott <[log in to unmask]>:
>
>> Dear James:
>>
>> It is extremely hard to make generalizations, but if it takes months for
>> crystals to appear, the following questions are worth asking:
>>
>> 1. Is the actual concentration of macromolecule, precipitating agent,
>> etc
>> required for crystallization higher than what you are using? A closed
>> vapor diffusion chamber is somewhat permeable to water vapor, and in the
>> course of five months, you could easily lose half of the water. Try
>> doubling the protein concentration, or crank up the concentration of one
>> or more components in the reservoir solution.
>>
>> 2. Is the actual macromolecule you crystallized the same as what you
>> started with? Slow degradation, or proteolysis, may be taking place,
>> and
>> the crystals you are growing are something else. (Similarly, if you
>> have
>> a cofactor or coenzyme, as these could slowly decompose into the
>> critical
>> ingredient for crystallization).
>>
>> 3. It might be too cold, and warming things up a bit might speed up
>> crystallization.
>>
>> Hopefully that will get you started before the experts reply.
>>
>> Bill
>>
>>
>>
>>
>>
>> On Sun, October 11, 2009 8:27 am, james09 pruza wrote:
>>> Dear crystallographers,
>>>
>>> Sorry for the non-ccp4 query. I am new to this field and need some
>>> suggestions. My question is, why some protein takes longer time to
>>> crystallize, say 6-8 months, and it is the only condition to get the
>>> crystals.? What are the ways to get the crystals faster.
>>>
>>> The crystal appears with 60% of 2-Methyl, 2-4 Pentane Diol and only at
>>> 4
>>> degree with very low concentration of NaCl. I have got some of the
>>> sugggestions earlier from the CCP4-discussion board for microseeding,
>>> but
>>> it
>>> did not work.
>>>
>>> All suggestions from the experts are welcome.
>>>
>>> Thanks.
>>> James
>>>
>>
>>
>>
>> William G. Scott
>>
>> Contact info:
>> http://chemistry.ucsc.edu/~wgscott/
>>
>
>
William G. Scott
Contact info:
http://chemistry.ucsc.edu/~wgscott/
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