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Questions and Answers
What is the primary purpose of electrophoresis in laboratory techniques?
What is the primary purpose of electrophoresis in laboratory techniques?
What is the function of the pores in the gel during electrophoresis?
What is the function of the pores in the gel during electrophoresis?
Why is electrophoresis important in protein research and genetic mutation research?
Why is electrophoresis important in protein research and genetic mutation research?
What happens to the smaller proteins during electrophoresis?
What happens to the smaller proteins during electrophoresis?
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Why is it not common to run a whole DNA molecule from a cell onto a gel?
Why is it not common to run a whole DNA molecule from a cell onto a gel?
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What is the function of ethidium bromide in the preparation of the gel?
What is the function of ethidium bromide in the preparation of the gel?
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What is the primary reason for chopping up DNA using inscription enzymes?
What is the primary reason for chopping up DNA using inscription enzymes?
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What is the purpose of the dye bromophenol blue in gel electrophoresis?
What is the purpose of the dye bromophenol blue in gel electrophoresis?
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In which direction do the DNA molecules migrate during gel electrophoresis?
In which direction do the DNA molecules migrate during gel electrophoresis?
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What is the typical voltage used per centimeter of gel during electrophoresis?
What is the typical voltage used per centimeter of gel during electrophoresis?
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Study Notes
Electrophoresis
- A laboratory technique used to separate DNA, RNA, or proteins based on their size and electrical charge.
- Agarose gel is commonly used for DNA separation, with a concentration of 0.8-2%.
- An electric current is used to move molecules through the gel, which acts as a sieve, separating molecules by size.
Importance of Electrophoresis
- Widely used in basic research for understanding gene and protein function.
- Essential in protein research and genetic mutation research, as mutated proteins or DNA often have altered sizes.
- Diagnostically important for detecting genetic mutations and forensic analysis.
Practical Technique
- Electrophoresis is typically performed in a box with a positive charge at one end and a negative charge at the other.
- Charged molecules migrate towards the opposite charge, with smaller molecules moving faster than larger ones.
- Proteins are usually analyzed by their entire length, with smaller proteins migrating farther into the gel and larger ones staying at the top.
DNA Analysis
- DNA is typically too large to run on a gel, so it's chopped into smaller pieces using inscription enzymes.
- These pieces are then separated based on their size, with smaller pieces migrating farther into the gel.
Preparation of Gel Electrophoresis
- Agarose is mixed with a buffer (TBE) and heated to ensure complete dissolution.
- Ethidium bromide is added to the same buffer used in the buffer tank.
- The sample (DNA) is prepared, mixed with a dye (e.g., bromophenol blue), and loaded into the gel.
- An electric current of 5-10 volts/cm is applied, causing DNA to migrate from negative to positive.
- The gel is then visualized under UV light.
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Description
A laboratory technique used to separate DNA, RNA, or protein molecules based on their size and electrical charge. Learn about agarose gel electrophoresis and how it works.
