Question

How can you isolate nuclei from cells?

Answer 1

The process of isolation of nuclei from the cell is called enucleation. The nucleus is a membrane-bound organelle that contains eukaryotic organisms' genetic material (DNA). It is the largest organelle that takes up about a tenth of the entire volume of cells inside the cell.

Complete answer:
The nucleus is a highly specialized organelle of the cell, complex and heterogeneous. It contains most of the hereditary information of the cell and controls most of the functions of the cell. The studies on gene transcription, RNA synthesis and processing, and posttranscriptional gene silencing are enhanced by isolated nuclei availability. The process of enucleation requires a microscope, a suction pipette and a glass needle. The process of enucleation is mostly done with oocytes or egg cells. In the first step to retain the egg cell, a suction pipette is used and a glass needle to remove the nucleus from the cell. In the second step the glass needle is pushed through the zona pellucida that surrounds the egg cell. With the help of a glass needle the nuclei is removed out of the nuclei.

Additional information:
A homogenization stage followed by centrifugation via high-density glycerol or sucrose was involved in the isolation of nuclei. Although this technique can be used to prepare clean nuclei with little cytoplasmic contamination, it is typically time consuming and can allow protein degradation, protein modification, and leaching of nucleus components to occur.

Note:
Isolated nuclei in higher plants have recently been used to investigate biochemical aspects. It is difficult to obtain adequate yields of intact nuclei from plant materials within a short period of time. Isolated nuclei are also used to obtain high-quality chromosomal DNA, to determine the level of ploidy and to measure nuclear DNA content using flow cytometry with minimized chloroplast and mitochondrial DNA contamination.