Hi,

 

I have a 22kDa protein that the floopy N & C terminus have been deleted. It was 
crystallized in 35%MPD/0.1m Tris (pH 8.5)/0.2M/(NH4)2SO4 in 2 - 3 days. 
Previously, a few small twining crystals were grown in this condition.

 

I tried Hampton Research 96-additive screen . Additives such as glycerol, 
dioxane, etc didn't work well to improve/reduce twining issue.

 

The 0.3% DMSO is the best additive I found that can grow a single crystal in 
the 1(pro)+0.8(buff)+0.2(add) drop.

 

However, If looking careful under microscope it may be some other crystals 
growing inside that single crystal(hardly see under the microscope, but I can 
see it in other drops). I conducted the x-ray diffraction experiment for this 
0.1x 0.1 mm crystal for which no cryoprotectant is required. The highest 
resolution is 2A but there are a lot of smear on the diffraction pattern.  
Thus, the index procedures failed due to the crystal quality.

 

Do you have any brilliant ideas to improve the crystal growing condition in my 
case in order to get a truly single crystal?

 

regards,

Heng 

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