molecular cloning

Examples of molecular cloning in the following topics:

• Hosts for Cloning Vectors

  • The majority of molecular cloning experiments begin with a laboratory strain of the bacterium E. coli (Escherichia coli) as the host.
  • A very large number of host organisms and molecular cloning vectors are in use, but the great majority of molecular cloning experiments begin with a laboratory strain of the bacterium E. coli (Escherichia coli) and a plasmid cloning vector.
  • If the DNA to be cloned is exceptionally large (hundreds of thousands to millions of base pairs), then a bacterial artificial chromosome or yeast artificial chromosome vector is often chosen.
  • In practice, however, specialized molecular cloning experiments usually begin with cloning into a bacterial plasmid, followed by subcloning into a specialized vector.

• Recombinant DNA Technology

  • Molecular cloning permits the replication of a specific DNA sequence in a living microorganism.
  • In standard molecular cloning experiments, the cloning of any DNA fragment essentially involves seven steps:
  • Although a very large number of host organisms and molecular cloning vectors are in use, the great majority of molecular cloning experiments begin with a laboratory strain of the bacterium E. coli (Escherichia coli) and a plasmid cloning vector.
  • Polymerase chain reaction (PCR) methods are often used for amplification of specific DNA or RNA (RT-PCR) sequences prior to molecular cloning.
  • The creation of recombinant DNA is in many ways the simplest step of the molecular cloning process.

• Molecular and Cellular Cloning

  • Molecular cloning reproduces the desired regions or fragments of a genome, enabling the manipulation and study of genes.
  • In general, the word "cloning" means the creation of a perfect replica; however, in biology, the re-creation of a whole organism is referred to as "reproductive cloning."
  • Long before attempts were made to clone an entire organism, researchers learned how to reproduce desired regions or fragments of the genome, a process that is referred to as molecular cloning.
  • Plasmids have been repurposed and engineered as vectors for molecular cloning and the large-scale production of important reagents such as insulin and human growth hormone.
  • This diagram shows the steps involved in molecular cloning, where regions or fragments of a genome are reproduced to allow the study or manipulation of genes and their protein products.

• Selection

  • Artificial selection is widely used in the field of microbial genetics, especially molecular cloning.
  • Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms.
  • Molecular cloning methods are central to many contemporary areas of modern biology and medicine.
  • In a conventional molecular cloning experiment, the DNA to be cloned is obtained from an organism of interest.
  • Molecular cloning is similar to polymerase chain reaction (PCR) in that it permits the replication of a specific DNA sequence.

• Obtaining DNA

  • Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms.
  • Molecular cloning generally uses DNA sequences from two different organisms: the species that is the source of the DNA to be cloned, and the species that will serve as the living host for replication of the recombinant DNA.
  • Although a very large number of host organisms and molecular cloning vectors are used, the great majority of molecular cloning experiments begin with a laboratory strain of the bacterium E. coli (Escherichia coli) and a plasmid cloning vector.
  • The creation of recombinant DNA is in many ways the simplest step of the molecular cloning process.
  • Explain the methods of obtaining DNA for molecular cloning experiments and the process of creating a recombinant DNA molecule

• Reproductive Cloning

  • Reproductive cloning, possible through artificially-induced asexual reproduction, is a method used to make a clone of an entire organism.
  • Reproductive cloning is a method used to make a clone or an identical copy of an entire multicellular organism.
  • This idea forms the basis for reproductive cloning.
  • It can be used for either therapeutic cloning or reproductive cloning.
  • Dolly the sheep was the first mammal to be cloned.

• Shuttle Vectors and Expression Vectors

  • Therefore, to make the purification process easy, the cloned gene should have a tag.
  • Expression vectors are used for molecular biology techniques such as site-directed mutagenesis.
  • In general, DNA vectors that are used in many molecular-biology gene-cloning experiments need not result in the expression of a protein.
  • The pGEX-3x plasmid is a popular cloning vector.
  • Please note the presence of a multiple cloning site, a promoter, a repressor, and a selectable marker.

• Vectors for Genomic Cloning and Sequencing

  • In molecular biology, a vector is a DNA molecule used as a vehicle to transfer foreign genetic material into another cell.
  • In molecular biology, a vector is a DNA molecule used as a vehicle to transfer foreign genetic material into another cell.
  • Modern plasmids generally have many more features, notably including a "multiple cloning site" which includes nucleotide overhangs for insertion of an insert, and multiple restriction enzyme consensus sites to either side of the insert.
  • The vectors can be extracted from the bacteria, and the multiple cloning sites can be cut by restriction enzymes to excise the hundredfold or thousandfold amplified insert.

• Genetic Analysis

  • Plasmid profiling using molecular, biochemical, and microbial techniques is essential to understanding the mechanism of pathogenicity and to fuel genetic engineering.
  • Many of the techniques used to study whole genomes are conventional molecular biology techniques adapted to operate effectively with DNA in a much larger size range.
  • PFGE is essential for estimating the sizes of whole genomes/chromosomes prior to sequencing and is necessary for preparing large DNA fragments for large insert DNA cloning and analysis of subsequent clones.
  • DNA cloning is another technique fundamental to molecular biology that requires adaptation in order to be useful in studying DNA at a whole genome scale.
  • Summarize the techniques used to study genomes: PFGE. ordered clone approach, direct shotgun sequencing and microarray hybridization

• Molecular Products from Microbes

  • The isolation of molecular products from microbes is considered to be a key component of molecular biology research.
  • The expansion and growing popularity of the field of molecular biology has resulted in a higher demand for tools used to study molecular biology.
  • The field of molecular biology specifically deals with the molecular mechanisms of a cell and focuses on the regulation of cellular interactions.
  • The following is a brief overview of some of the molecular products derived from microbes that allow for the performance of popular molecular biology techniques.
  • The isolation of approximately 3000 restriction enzymes has allowed molecular biologists to utilize them in processes such as cloning and the production of recombinant DNA .