Setting up hanging drop protein crystals

12 Aug

Setting up hanging drop protein crystals

Monday, August 12, 2013

VtS-PCS-3

Today VTS learned how to set-up hanging-drop vapor diffusion protein crystals using the VDX pre-greased plates. Over the last four to six weeks, VTS has been learning how to manually grease VDX plates for setting up crystals of RNA+Protein complexes in one of the structural biology labs. However, since the RNA+Protein complexes are very expensive and since VTS has been only volunteering his work was limited to preparing the trays.

VtS-PCS-1

However, today he used the lysozyme solutions left-over from another experiment to set-up the hanging drop vapor diffusion protein crystallization trays.

He used 3 microliter of 30 milligram/milliliter protein and mixed it with 3 microliters of 10 % (w/v) sodium chloride solution in sodium acetate buffer, pH 4.8. He then mixed these solutions on top of siliconized cover slip. He added 600 microliters of 10 % (w/v) sodium chloride solution in sodium acetate buffer, pH 4.8 to the well. He then covered the well with the inverted coverslip containing the protein+precipitant mixture.

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