Methods of DNA Cloning

There are several types of DNA cloning methods, each with its specific applications and advantages. Here are some common types of DNA cloning:

Traditional Method

In the traditional cloning method, DNA fragments containing the gene of interest and a cloning vector are cut with the same restriction enzyme(s) to create complementary ends.The ends of the DNA fragment and vector are ligated together using DNA ligase, forming recombinant DNA molecules. The recombinant DNA is then introduced into host cells, where it is replicated to produce multiple copies of the cloned DNA fragment.

PCR Cloning

PCR cloning involves the use of polymerase chain reaction (PCR) to amplify the gene of interest from genomic DNA or cDNA. Primers used in PCR contain sequences that are complementary to the ends of the target gene and the cloning vector. PCR products are then ligated into a cloning vector and introduced into host cells for propagation.

Ligation Independent Cloning

Ligation-independent cloning (LIC) is a method that utilizes single-stranded overhangs to ligate DNA fragments. PCR primers are designed to add single-stranded overhangs to the ends of DNA fragments. These overhangs anneal to complementary sequences in the cloning vector, allowing for the formation of recombinant DNA molecules without the need for restriction enzymes or DNA ligase.

Seamless Cloning

Seamless cloning methods, such as Gibson Assembly or In-Fusion cloning, enable the joining of DNA fragments with overlapping ends without the use of restriction enzymes or DNA ligase. DNA fragments with overlapping sequences are mixed with DNA polymerase, exonuclease, and DNA ligase enzymes to create seamless recombinant DNA molecules.

Recombinational Cloning

Recombinational cloning methods, such as Gateway cloning, utilize site-specific recombination systems to facilitate the transfer of DNA fragments between different vectors. DNA fragments are first cloned into a donor vector, generating an entry clone. The entry clone is then recombined into a destination vector, allowing for the transfer of the DNA fragment between vectors without the need for restriction enzymes or DNA ligase.

Also Read: Cloning Vectors – CBSE Notes Class 12

DNA Cloning – An Overview

DNA cloning is an essential technique in molecular biology that enables researchers to produce multiple copies of a specific DNA sequence. It involves isolating a target DNA fragment and inserting it into a vector (such as a plasmid or a viral genome), which is then introduced into a host organism (usually bacteria or yeast) to produce multiple copies of the inserted DNA. In this article, find the DNA cloning notes and diagram.

Table of Content

DNA Cloning Definition
What is DNA Cloning?
Diagram of DNA Cloning
Methods of DNA Cloning
Steps of DNA Cloning
Application of Gene Cloning
DNA Cloning Vector