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Q.How can we make bacterial cell competent for transformation with recombinant DNA ?

Kerala DhseKerala DHSE Plus Two Board 2023Subjective· 2mImportance★★★★★
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Bacterial cells are made competent to take up recombinant DNA by treating them with a specific concentration of a divalent cation such as calcium chloride, which enlarges pores in the cell wall/membrane, and then giving the DNA-cell mixture a brief heat shock to encourage DNA entry.

Bacteria do not readily take up foreign DNA on their own — since most bacterial cells lack a natural mechanism for DNA uptake, they must be made 'competent' first:

  1. 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 cell by making the cell wall porous.
  2. The recombinant DNA is then incubated with these competent cells on ice.
  3. The mixture is then given a brief heat shock (typically 42°C for about 90 seconds), after which it is placed back on ice. …

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