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Increase the concentration of ammonium sulfate (cryosalt) (or primary salt acting as a precipitant) in increments of 10% in the presence of 5 to 10% v/v Glycerol.
Evaluate 25-30% w/v glucose, or trehalose or sucrose as cryoprotectants for crystals grown in Ammonium sulfate. Acta Cryst. (2002). D58, 1664-1669. Crystallization of RNA/protein complexes. M. Garber, G. Gongadze, V. Meshcheryakov, O. Nikonov, A. Nikulin, A. Perederina, W. Piendl, A. Serganov and S. Tishchenko.
15 to 30% v/v Ethylene glycol, DMSO, or Glycerol.
Try 1 – 2 M Sodium malonate as a cryoprotectant. Acta Cryst. (2003). D59, 2356-2358. Malonate: a versatile cryoprotectant and stabilizing solution for salt-grown macromolecular crystals. T. Holyoak, T. D. Fenn, M. A. Wilson, A. G. Moulin, D. Ringe and G. A. Petsko.
Place a drop of 75% v/v Paratone-N, 25% v/v Paraffin Oil next to the drop containing the crystal. Remove the crystal from the drop using a CryoLoop and dip the crystal into the oil. Keeping the CryoLoop and crystal immersed, gently move the CryoLoop containing the crystal from the oil into the reagent, back into the oil, back into the reagent, repeating this several times to remove some of the aqueous (reagent) layer from the crystal surface. Cryo cool the crystal.
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