electrophoresis
Microbiology
Biology
Examples of electrophoresis in the following topics:
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Northern Blots
- Northern blotting involves the use of electrophoresis to separate RNA samples by size and detection with a hybridization probe complementary to part of or the entire target sequence.
- The term 'Northern blot' actually refers specifically to the capillary transfer of RNA from the electrophoresis gel to the blotting membrane.
- RNA samples are then separated by gel electrophoresis.
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Basic Techniques to Manipulate Genetic Material (DNA and RNA)
- Basic techniques used in genetic material manipulation include extraction, gel electrophoresis, PCR, and blotting methods.
- Gel electrophoresis is a technique used to separate molecules on the basis of size using this charge and may be separated as whole chromosomes or fragments.
- Gel electrophoresis separates the nucleic acid fragments according to their size.
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Immunoblot Procedures
- Proteins are generally separated by size using sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis.
- In clinical diagnostic settings, immunoelectrophoresis is applied, which involves the electrophoresis of serum or urine followed by immunodiffusion.
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DNA Sequencing Based on Sanger Dideoxynucleotides
- Following rounds of template DNA extension from the bound primer, the resulting DNA fragments are heat denatured and separated by size using gel electrophoresis.
- DNA sequencers carry out capillary electrophoresis for size separation, detection and recording of dye fluorescence, and data output as fluorescent peak trace chromatograms.
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Genetic Analysis
- Pulsed field gel electrophoresis (PFGE) is an adaptation of conventional agarose gel electrophoresis that allows extremely large DNA fragments to be resolved (up to megabase size fragments).
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Western Blots
- The technique uses gel electrophoresis to separate native proteins by 3-D structure or denatured proteins by the length of the polypeptide.
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DNA Protection Analysis
- The method uses an enzyme, deoxyribonuclease (DNase, for short) to cut the radioactively end-labeled DNA, followed by gel electrophoresis to detect the resulting cleavage pattern.
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Sickle-Cell Disease
- Abnormal hemoglobin forms can also be detected on hemoglobin electrophoresis, a form of gel electrophoresis on which the various types of hemoglobin move at varying speeds.
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Basic Techniques in Protein Analysis
- The western blot, or protein immunoblot, is a technique that combines protein electrophoresis and antibodies to detect proteins in a sample.
- The protein sample is first separated by gel electrophoresis, then transferred to a nitrocellulose or other type of membrane, and finally stained with a primary antibody that specifically binds the protein of interest.
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Multiplex and Real-Time PCR
- It is confirmed by agarose gel electrophoresis for qualitative results.
- The results from multiplex PCR can be analyzed using gel electrophoresis or using fluorophores for analysis using during the reaction.