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Q.Write the steps in making the bacterial cell competent to take up the recombinant DNA.
Kerala DhseKerala DHSE Plus Two Board 2025Subjective· 2mImportance★★★★★
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Start your 14-day free trial to unlock the full solution →Bacteria do not readily take up foreign DNA on their own, so they are chemically treated to make their cell walls temporarily permeable ('competent') to DNA, then given a controlled temperature shock that drives the DNA inside.
Steps to make bacterial cells competent to take up recombinant DNA:
- Divalent cation treatment: the bacterial cells are treated with a specific concentration of a divalent cation, most commonly calcium chloride (CaCl2). This increases the efficiency with which DNA can enter the bacterium through pores in its cell wall.
- Incubation with recombinant DNA on ice: the treated (now 'competent') bacterial cells are mixed with the recombinant DNA and kept on ice for a period of time.
- Heat shock: the mixture is then briefly placed at 42°C (heat shock) for about 90 seconds.
- Return to ice: the cells are immediately placed back on ice. …
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