molecular cloning

(noun)

a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms.

Related Terms

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.

  • 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

  • Detecting Uncultured Microorganisms

    • However, recovery of DNA sequences longer than a few thousand base pairs from environmental samples was very difficult until recent advances in molecular biological techniques.
    • More specifically, the construction of libraries in bacterial artificial chromosomes (BACs) provided better vectors for molecular cloning.
    • Shotgun sequencing and screens of clone libraries reveal genes present in environmental samples.
    • (A) sampling from habitat; (B) filtering particles, typically by size; (C) Lysis and DNA extraction; (D) cloning and library construction; (E) sequencing the clones; (F) sequence assembly into contigs and scaffolds

  • 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 .

  • Plasmids as Cloning Vectors

    • Plasmids can be used as cloning vectors, allowing the insertion of exogenous DNA into a bacterial target.
    • In molecular biology, a vector is a DNA molecule used as a vehicle to transfer foreign genetic material into another cell.
    • All engineered vectors have an origin of replication, a multi-cloning site, and a selectable marker.
    • Modern plasmids generally have many more features, notably a "multiple cloning site"—with nucleotide overhangs for insertion of an insert—and multiple restriction enzyme consensus sites on either side of the insert.
    • The pGEX-3x plasmid is a popular cloning vector.