Selecting transformed cells from "summary" of Molecular Cloning by Joseph Sambrook,David William Russell
After the introduction of the foreign DNA into bacterial cells, the next crucial step in molecular cloning is the selection of cells that have taken up the desired DNA. This selection process is essential to identify and isolate the transformed cells that contain the recombinant DNA of interest. Without proper selection, it would be nearly impossible to distinguish between transformed and non-transformed cells, making the isolation of the desired DNA a challenging task.
To achieve this selection, scientists often use a technique known as antibiotic selection. In this method, the foreign DNA is usually introduced into the bacterial cells along with a gene that confers resistance to a specific antibiotic. This antibiotic resistance gene is typically located within the same vector as the foreign DNA, allowing for both the insertion of the desired DNA and the selection of transformed cells in a single step.
After the transformation process, the bacterial cells are plated onto agar plates containing the specific antibiotic to which the resistance gene confers protection. Only the transformed cells that have taken up the recombinant DNA, along with the antibiotic resistance gene, will be able to survive and grow on the antibiotic-containing plates. Non-transformed cells or those that have not taken up the foreign DNA will die in the presence of the antibiotic, allowing for the selective growth of transformed cells.
This antibiotic selection process is a powerful tool in molecular cloning as it enables researchers to easily identify and isolate the transformed cells that contain the desired DNA. By using antibiotics as a selective agent, scientists can efficiently screen large populations of bacterial cells and isolate those that have successfully incorporated the foreign DNA. This method has revolutionized the field of molecular biology, allowing for the rapid and efficient cloning of genes and other DNA sequences of interest.
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