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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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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:

  1. 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.
  2. 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.
  3. Heat shock: the mixture is then briefly placed at 42°C (heat shock) for about 90 seconds.
  4. Return to ice: the cells are immediately placed back on ice. …

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