Denaturation of DNA:In the denaturation process, the hydrogen bonds between

two strands are broken giving rise to two single strands. The covalent bonds of DNA remain
unaffected.

Denaturation of DNA double helix takes place by the

following denaturating agents:

(i) Denaturation by Temperature:

If a DNA solution is heated to approximately 90°C or above there will

be enough kinetic energy to denature the DNA completely causing it to
separate into single strands. This denaturation is very abrupt and is
accelerated by chemical reagents like urea and formamide.The
chemicals enhance the aqueous solubility of the purine and pyrimidine
groups. This separation of double helix is called melting as it occurs
abruptly at a certain characteristic temperature called denaturation
temperature or melting temperature (Tm).It is defined as temperature
at which 50% of the DNA is melted.If several samples of DNA are
melted, it is found that the Tm is highest for those DNAs that contain
the highest proportion of G—C. Actually the value is used to estimate
the percentage of G—C in a DNA sample. In fact, the Tm of DNA from
many species varies linearly with G—C content.

This relationship between Tm and G—C content arises due to guanine

and cytosine form three hydrogen bonds when base paired, whereas
adenine and thymine form only two.

DNA Denaturation through NaOH Treatment

Apart from heating, chemical denaturation can also be achieved through the use of NaOH. A certain
concentration of NaOH can be used to denature DNA safely, often in as little as one minute. As the
concentration of NaOH used is reduced, the process will take longer -- but the DNA can still be fully
denatured.

DNA Denaturation through Salt .

A high concentration of salt will cause DNA to naturally denature, given the right concentration of
salt. DNA denaturation with salts are similar to denaturation through the use of organic solvents. In
general, DNA denaturation through salt cannot be renatured.

Effect of denaturation of DNA:

● Increased absorption of UV light at 260nm wavelengths. The rate of absorption is directly

proportional to the rate of denaturation
● Viscosity decreases, which reflects the physical change occurred in the DNA structure

What is Renaturation?
Renaturation is also known as annealing. When the temperature and pH return to optimum biological
level, the unwound strand of DNA rewind and give back the dsDNA.

If the DNA is not completely denatured, the renaturation process is fast and a one-step process, but
if the DNAs are completely denatured then the renaturation process occurs in a two-step process.
First complementary strands come together by random collision and then rewinding takes place
forming a double helix.

Renaturation occurs when the denatured DNAs are cooled in suitable conditions. Renaturation also
depends on temperature, pH, length and constituents of the DNA structure. The renaturation rate is
directly proportional to the number of complementary sequences present.

With renaturation, absorption of UV (260nm) decreases and viscosity increases again.