Describe how, using mice, a researcher could carry out (1) organismalcloning, (2) production of ES cells, and (3) generation of iPS cells,focusing on how the cells are reprogrammed. (The procedures arebasically the same in humans and mice.)
Molecular Techniques
Molecular techniques are methods employed in molecular biology, genetics, biochemistry, and biophysics to manipulate and analyze nucleic acids (deoxyribonucleic acid (DNA) and ribonucleic acid (RNA)), protein, and lipids. Techniques in molecular biology are employed to investigate the molecular basis for biological activity. These techniques are used to analyze cellular properties, structures, and chemical reactions, with a focus on how certain molecules regulate cellular reactions and growth.
DNA Fingerprinting and Gel Electrophoresis
The genetic makeup of living organisms is shown by a technique known as DNA fingerprinting. The difference is the satellite region of DNA is shown by this process. Alex Jeffreys has invented the process of DNA fingerprinting in 1985. Any biological samples such as blood, hair, saliva, semen can be used for DNA fingerprinting. DNA fingerprinting is also known as DNA profiling or molecular fingerprinting.
Molecular Markers
A known DNA sequence or gene sequence is present on a chromosome, and it is associated with a specific trait or character. It is mainly used as a genetic marker of the molecular marker. The first genetic map was done in a fruit fly, using genes as the first marker. In two categories, molecular markers are classified, classical marker and a DNA marker. A molecular marker is also known as a genetic marker.
DNA Sequencing
The most important feature of DNA (deoxyribonucleic acid) molecules are nucleotide sequences and the identification of genes and their activities. This the reason why scientists have been working to determine the sequences of pieces of DNA covered under the genomic field. The primary objective of the Human Genome Project was to determine the nucleotide sequence of the entire human nuclear genome. DNA sequencing selectively eliminates the introns leading to only exome sequencing that allows proteins coding.
Describe how, using mice, a researcher could carry out (1) organismal
cloning, (2) production of ES cells, and (3) generation of iPS cells,
focusing on how the cells are reprogrammed. (The procedures are
basically the same in humans and mice.)
Cloning is the method by which an organism is created by the cells derived from a parent organism. The cloned organism is genetically identical to the parent organism. The DNA (deoxyribonucleic acid) sequence of the cloned organism is the same as that of parent organisms. The first animal to be cloned was a sheep named Dolly at Roslin Institute, Scotland.
The cloning of mice can be carried out similarly as sheep cloning. The method used for cloning is reproductive cloning. The somatic cells of a donor mice are taken and inoculated in a culture medium. The formulation of the medium is designed specifically to keep the somatic cells in a non-dividing resting state. An oocyte from a different strain of mice is taken. This oocyte is arrested at the metaphase II stage of meiosis. The next step is the enucleation or removal of the nucleus from the oocyte. The fusion of the enucleated oocyte and the somatic cells is triggered by bringing cells to close together and the application of the electrical pulse. The electrical pulse causes the destabilization of the cell membrane allowing them to fuse. The fused cell is placed in a culture medium to allow its growth to the blastocyst stage of embryonic development. The blastocyst is then implanted in the uterus of a surrogate mice to develop into an organism genetically identical to the donor mice.
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