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Proteins and proteomics
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2018
The western blot (alternatively, protein immunoblot) is an analytical technique used to detect specific proteins in a given sample of tissue homogenate or extract. It uses gel electrophoresis to separate native or denatured proteins by the length of the polypeptide (denaturing conditions) or by the 3D structure of the protein (native/non-denaturing conditions). The proteins are then transferred to a membrane (typically nitrocellulose or INDF), where they are probed (detected) using antibodies specific to the target protein. Now many reagent companies specialize in providing antibodies (both monoclonal and polyclonal antibodies) against tens of thousands of different proteins. Conunercial antibodies can be expensive, although the unbound antibody can be reused between experiments. This method is used in the fields of molecular biology, biochemistry, immunogenetics, and other molecular biology disciplines. The method originated from the laboratory of George Stark at Stanford. The name western blot was given to the technique by W. Neal Burnette and is a play on the name Southern blot, a technique for DNA detection developed earlier by Edwin Southern (Figure 3.15).
Glossary of scientific and technical terms in bioengineering and biological engineering
Published in Megh R. Goyal, Scientific and Technical Terms in Bioengineering and Biological Engineering, 2018
Gel electrophoresis is a method for separation and analysis of macromolecules (DNA, RNA and proteins) and their fragments, based on their size and charge. Gel electrophoresis uses a gel as an anticonvective medium and/or sieving medium during electrophoresis, the movement of a charged particle in an electrical field. Gels suppress the thermal convection caused by application of the electric field, and can also act as a sieving medium, retarding the passage of molecules; gels can also simply serve to maintain the finished separation, so that a post electrophoresis stain can be applied.
Approaches to the Measurement of Biological Pollutants
Published in Somenath Mitra, Pradyot Patnaik, Barbara B. Kebbekus, Environmental Chemical Analysis, 2018
Somenath Mitra, Pradyot Patnaik, Barbara B. Kebbekus
Gel electrophoresis used for the separation of DNA, RNA, and protein molecules using an electric field applied to a gel matrix. Gel electrophoresis is often the analytical step that follows the PCR amplification of DNA. However, it can also be used as a sample preparation technique for separation of specific species to be later analyzed via other methods such as mass spectrometry, fluorescence based methods, Southern blotting, and even PCR. A typical gel electrophoresis setup is shown in Figure 10.10.
Evolutionary DNA Computing Algorithm for Job Scheduling Problem
Published in IETE Journal of Research, 2018
Gudar J. Ibrahim, Tarik A. Rashid, Ahmed T. Sadiq
For that reason, a Watson–Crick complementary string can be produced via a Turing machine which can code various programs to emulate games such as chess, etc. The basic DNA computing operations that are widely used can be described as follows [23,34,36,38,41–49]: Pairing of Watson–Crick: this prevalent, clearly, strands of DNA can have their Watson–Crick string complements. If a DNA molecule comes across Watson–Crick original complementary strand, the process of pairing will take place. Therefore, both DNA strands can get joined and annealed to yield the double helix.Polymerases: DNA polymerases can generate DNA complementary strands to Watson–Crick DNA original string. Thus, information can be copied from a specific molecule to another one via polymerases.Ligases: these are very useful for connecting macules. The process of ligase would use two strands of DNA to generate one.Nucleases: collapse and thus suppress DNA and RNA.Gel electrophoresis: they are used to examine and set apart protein macromolecules, DNA, and RNA.Synthesis: basically, the DNA sequences are written on a piece of paper and sent to the facility of synthesis. Then, over 1000 of molecules of DNA contained in a tube are returned by the facility of synthesis within a couple of days.