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Explain the principle and any two uses of each of the following biochemical techniques :

(i) Partition Chromatography . 

(ii) X-Ray Crystallography.

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i) Partition Chromatography: Partition chromatography is the process of separation whereby the components of the mixture get distributed into two liquid phases due to differences in partition coefficients during the flow of mobile phase in the chromatography column. 

Here the molecules get preferential separation in between two phases, i.e., both stationary phase and mobile phase are liquid in nature. So molecules get dispersed into either phases preferentially. Polar molecules get partitioned into polar phase and vice-versa. This mode of partition chromatography applies to liquid-liquid, liquid-gas chromatography and not to solid-gas chromatography. Because partition is the phenomenon in between a liquid and liquid or liquid and gas or gas and gas. But not in solid involvement. Uses: 

• It is used to separate the amino acids. 

• It is extensively used for the study of lipid-soluble substances. 

(ii) The technique of single-crystal X-ray crystallography has three basic steps : The first and often most difficult step is to obtain an adequate crystal of the material under study. The crystal should be sufficiently large (typically larger than 100 microns in all dimensions), pure in composition and regular in structure. 

In the second step, the crystal is placed in an intense beam of X-rays, usually of a . single wavelength (monochromatic X-rays), producing the regular pattern of reflections. 

As the crystal is gradually rotated, previous reflections disappear and new ones appear; the intensity of every spot is recorded at every orientation of the crystal. Multiple data sets may have to be collected, with each set covering slightly more than half a lull rotation of the crystal and typically containing tens of thousands of reflection intensities. 

In the third step, these data combined computationally with complementary chemical information to produce and refine a model of the arrangement of atoms within the crystal. The final, refined model of the atomic arrangement now called a crystal structure is usually stored in a public database.

Uses: 

• It is used to determine the atomic and molecular structure of a crystal. 

• It is also used in X-ray diffraction to rotate the samples.

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