Question

Question: Summarize the steps required to amplify a given segment of DNA in vivo and in vitro.

Short answer

Answer

DNA amplification of a given segment occurs in vivo and in vitro through DNA replication and PCR reaction. Three main stages are required for in-vitro DNA amplification: denaturation, annealing, and extension.

In-vivo DNA amplification, on the other hand, happens during the cell cycle's S phase. Certain enzymes work on the parent DNA molecule to reveal its nitrogenous bases, allowing the daughter DNA molecule to be formed before the expansion of DNA base pairing.

Step-by-step solution

DNA amplication

Multiple copies of a DNA sequence can be created via DNA amplification.In Vivo, DNA amplification occurs when DNA is amplified inside the body. In vitro amplification of DNA is a laboratory method for amplifying DNA molecules.

DNA replication in vivo

The nucleus of the daughter cell generated during cell division has the identical copy of genes as the mother cell. There are several enzymes that aid in the in-vivo process.

DNA helicase, polymerase, RNA primase, and DNA ligase are enzymes involved in DNA replication.The DNA strand is denatured, allowing the nitrogenous bases to be exposed. Following that, the matching of these nitrogenous bases begins, synthesizing a new strand of DNA.

DNA amplification in vitro

In-vitro DNA amplification is a method that creates a new DNA strand in the lab. The polymerase chain reaction (PCR) technology is commonly used in vitro. DNA replication PCR, like in-vivo, necessitates the use of enzymes. Taqpolymerase is a DNA polymerase which is extremely stable at high temperatures, and at 70 degrees Celsius, it becomes extremely active.This is why, in in-vitro circumstances, this is the principal enzyme employed for DNA amplification; high temperatures are required to denature the template DNA to separate its strands repeatedly.

Primers are another component of the PCR process. Primers are the molecules that start DNA synthesis. These are single-stranded, short DNA strands. The primer is attached to its template DNA using the base pairing approach.

Polymerase Chain Reaction

PCR occurs in 3 steps:

• Denaturation: Denaturation is the process of uncoiling DNA to create a single strand that can be used as a template by heating it to 96 degrees Celsius.
• Annealing: The primers attach to the template DNA when the temperature is lowered to 50 to 55 degrees Celsius.
• Extension: The last stage is an extension, in which the increasing temperature aids the action of the enzyme DNA polymerase, allowing it to begin base pairing primers with complement DNA.