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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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Start your 14-day free trial to unlock the full solution →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:
- 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.
- The recombinant DNA is then incubated with these competent cells on ice.
- 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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