Concept explainers
To determine: The reason why the renaturation of E. coli DNA is a monophasic process, while the renaturation of human DNA is biphasic.
Introduction: Denaturation of DNA is a mono-molecular reaction and includes melting of DNA into two complementary strands. Renaturation of DNA is a bi-molecular reaction because it depends on the concentration of two complementary strands. It is based on the concentration of DNA (Co) and the time taken for renaturation (t).
Explanation of Solution
Explanation:
The percentage of single-stranded DNA is plotted as a function of Cot; that is, the initial concentration of DNA with time. The midpoint of the curve provides Cot1/2; that is, the time required for half renaturation. The genome of E. coli consists of approximately 50, 00,000 bp of unique sequences that are present as single copies. After plotting the percentage of single-stranded DNA as a function of Cot, a single sigmoidal curve is observed. This is because each denatured fragment would reassociate with its complementary strand. This concludes that renaturation of E. coli DNA as a monophasic process. However, in contrast, the human genome is complex as it has repetitive sequences. After denaturation, DNA fragments find their complementary sequences as the first phase. In the second phase, they renature to form double-stranded DNA strands resulting in the formation of a biphasic renaturation curve.
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