Step I: Homogenization 1. Take g onion and make slices of 1-2 mm pieces 2. Obtai

Question

Step I: Homogenization 1. Take g onion and make slices of 1-2 mm pieces 2. Obtain a mass of 20 3. Add 100 ml homogenizing solution. gresh sliced onion in a 250 ml conical flask. 4. Mix thoroughly and incubate the flask at 65 C for 15 minutes What happens at this stage by: Heat (63 C): soften onion tissues, loosen cells, denature enzymes. SDS: dissolves membranes, and denatures protein. 5. Incubate the flask in ice bath (pieces of ice in water) to lower temperature to 20°C. 6. Pour cooled onion slice preparation in a blender and fasten the lid. Homogenize for 30 seconds at low speed, followed by 30 seconds at high speed. What happens by: Homogenization: Na-citrate: 7. Pour homogenate from the blender and keep on ice for 10 minutes. 25

Explanation / Answer

Homogenization is a method which is used to disrupt the cell tissue and lyse the cell. With the help of homogenization step cells are separated and breaks them open. Homogenization is done with a mechanical instruments such as blender. Here the process of homogenization breaks down the cell wall membrane and nuclear membrane of the onion sample which will allow the organales to release out.

Na-citrate buffer is used to stabilize the DNA of the cells by making a Na+ shell around the negative charged phosphate group of DNA. On the other hand citrate inactivates the DNAse which breaks down the DNA.

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