The technology used for producing artificial DNA through the combination of different genetic materials (DNA) from different sources is referred to as recombinant DNA technology. Recombinant DNA technology is popularly known as genetic engineering. It involves the selection of the desired gene for administration into the host followed by a selection of the perfect vector with which the gene has to be integrated and recombinant DNA formed.
Inserting the desired gene into the genome involves the selection of the desired gene for administration into the host followed by a selection of the perfect vector with which the gene has to be integrated and recombinant DNA formed. Thus the recombinant DNA has to be introduced into the host. And at last, it has to be maintained in the host and carried forward to the offspring.
Procedure
-
Isolation of genetic material
A DNA sequence is the sequence of nucleotides in a DNA molecule. It is written as a succession of letters representing the primary structure of a DNA molecule or strand. -
Cutting the gene at the recognition sites
The restriction enzymes play a major role in determining the location at which the desired gene is inserted into the vector genome. These reactions are called 'restriction enzyme digestions'. -
Amplifying the gene copies through polymerase chain reaction (PCR)
It is a process to amplify a single copy of DNA into thousands to millions of copies once the proper gene of interest has been cut using restriction enzymes. -
Ligation of DNA molecules
In this step of ligation, the joining of the two pieces - a cut fragment of DNA and the vector together with the help of the enzyme DNA ligase. -
Insertion of recombinant DNA into host
In this step, the recombinant DNA is introduced into a recipient host cell. This process is termed as transformation. Once the recombinant DNA is inserted into the host cell, it gets multiplied and is expressed in the form of the manufactured protein under optimal conditions.
Fig. 1 The basic flow chart for recombinant DNA technology.
Tools
- Enzymes: the restriction enzymes help to cut, the polymerases help to synthesize and the ligases help to bind.
- Vectors: help in carrying and integrating the desired gene. Plasmids and bacteriophages are the most common vectors in recombinant DNA technology that are used as they have a very high copy number.
- Host organism: into which the recombinant DNA is introduced. The host is the ultimate tool of recombinant DNA technology which takes in the vector engineered with the desired DNA with the help of the enzymes.
Applications
- Detection of the presence of HIV in a person
- Gene Therapy: it is used as an attempt to correct the gene defects which give rise to heredity diseases
- Clinical diagnosis: ELISA is an example of application of recombinant
- Production of genetically-modified organisms in agriculture, such as Flavr Savr tomatoes, golden rice rich in proteins, and Bt-cotton to protect the plant against ball worms and a lot more
- For the production of insulin in the field of medicines
Related Products
Thermal Cycler
Thermal Cycler is a laboratory apparatus that amplify segments of DNA using PCR. A thermal cycler is also commonly called a DNA Amplifier, PCR Machine, or Thermocycler. The cycle normally lowers or raises the block's temperature in preprogrammed discrete steps. A thermal cycler is important for a laboratory dealing in molecular biology and gene cloning.
Digital PCR System (dPCR)
Digital PCR Systems (dPCR) is incredibly useful in applications such as mutation detection, copy number variation, rare sequence detection, gene expression analysis, miRNA analysis and next generation sequencing sample quantification. It provides a precise measure of DNA molecules in each drop. In some cases, Digital PCR Systems (dPCR) must be used in combination with a 96-deep well reaction module.
Real-time PCR System
Real-time PCR System can quantify DNA copies and enable experiments in gene expression, genetic variation, genotyping, and specific detection of rare targets, bacteria, and viruses. It measures signals generated by fluorescent probes that are proportional to DNA amplification, allowing accurate quantification. It is capable of quantifying very small amounts of DNA with good dynamic range.
Laboratory Incubator
An incubator is a device used to grow and maintain microbiological cultures or cell cultures. It is made up of a chamber with a regulated temperature. An incubator maintains optimal temperature, humidity, and other conditions such as the CO2 and oxygen content of the atmosphere inside. Incubators are essential for much experimental work in cell biology, microbiology, and molecular biology.
Electrophoresis System
Electrophoresis equipment applies an electric charge to molecules, causing them to migrate towards their oppositely charged electrode. The technique is found in all research and clinical laboratories utilizing DNA and protein applications, and is divided into gel and capillary techniques. Choice of electrophoresis equipment depends on the molecule of study, appropriate method of separation and downstream application.
Digital Gel Imaging System
Gel imaging or gel documentation (gel doc) systems are used for the analysis of proteins, antibodies and nucleic acid immobilized in polyacrylamide or agarose gels, membranes or microarrays. Systems come in a variety of configurations depending on throughput and sample type. It is widely used in: monoclonal and polyclonal antibody binding affinities, gel and blot imaging, colony counting, immunoassay, multiplex protein detection and other fields.
STEMart provides you with a variety of recombinant DNA technology equipment or consumables to meet your various R&D and application needs. If you have any questions or requirements for recombinant DNA technology, please feel free to contact us.