Analytical Chemistry and Separation Techniques

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Questions and Answers

What is a common use of gel electrophoresis?

To increase DNA replication

To synthesize proteins

To separate DNA fragments (correct)

To separate RNA fragments

Which gel type offers higher resolution than agarose gel for small DNA fragments?

Starch gel

Cellulose gel

Polyacrylamide gel (correct)

Gelatin gel

Why is polyacrylamide gel electrophoresis used instead of agarose gel for small DNA fragments?

It is less expensive

It takes less time

It provides higher resolution (correct)

It is easier to prepare

What factor primarily affects the movement of DNA in gel electrophoresis?

Temperature of the gel Signup and view all the answers

What is the charge of DNA fragments during gel electrophoresis?

Negative Signup and view all the answers

In gel electrophoresis, DNA moves towards which electrode?

Cathode Signup and view all the answers

What property of DNA allows it to be separated by gel electrophoresis?

Negative charge Signup and view all the answers

What is the function of the buffer solution in gel electrophoresis?

To provide a medium for electrical conductivity Signup and view all the answers

Which of the following staining methods is commonly used to visualize DNA in a gel after electrophoresis?

Ethidium bromide Signup and view all the answers

What is the role of the loading dye in gel electrophoresis?

To make the DNA visible under UV light Signup and view all the answers

Which of the following is NOT a type of high performance liquid chromatography column?

Solid Phase Signup and view all the answers

What does the stationary phase do in high performance liquid chromatography?

Separates the sample components based on interactions Signup and view all the answers

Which detector is commonly used in high performance liquid chromatography for compounds that absorb UV or visible light?

Diode array detector Signup and view all the answers

What is the primary purpose of gel electrophoresis in molecular biology?

To separate DNA fragments by size Signup and view all the answers

Which of the following gels is most commonly used for DNA separation?

Agarose Signup and view all the answers

Which of the following is a primary advantage of using agarose gel in electrophoresis?

Large pore size suitable for DNA Signup and view all the answers

In SDS-PAGE, what is the function of SDS (sodium dodecyl sulfate)?

To give them a uniform negative charge Signup and view all the answers

What does the term 'resolution' refer to in gel electrophoresis?

The ability to distinguish between two closely spaced DNA fragments Signup and view all the answers

Why is it important to use a molecular weight marker in protein gel electrophoresis?

To determine the molecular weight of proteins Signup and view all the answers

What type of gel electrophoresis is typically used for RNA separation?

Agarose gel electrophoresis with formaldehyde Signup and view all the answers

How can the accuracy of gel electrophoresis results be verified?

By repeating the experiment multiple times Signup and view all the answers

How can the resolution of gel electrophoresis be improved?

By using a different type of gel Signup and view all the answers

What is the purpose of using a comb during the preparation of a gel for electrophoresis?

To create wells for loading samples Signup and view all the answers

Which of the following is a common use of gel electrophoresis in medical diagnostics?

Detecting genetic mutations Signup and view all the answers

Which of the following is an example of adsorption chromatography?

Chromatography using a column containing silica gel or alumina Signup and view all the answers

What are the two main components required for all chromatographic methods?

Stationary phase and mobile phase Signup and view all the answers

Which of the following statements best describes the role of a gel in electrophoresis?

It acts as a stationary phase Signup and view all the answers

Study Notes

High Performance Liquid Chromatography (HPLC)

Solid Phase is not a type of HPLC column.
The stationary phase separates sample components based on interactions.

HPLC Detectors

Diode array detector is commonly used for compounds that absorb UV or visible light.

Gel Electrophoresis

Primary purpose: to separate DNA fragments by size.
Agarose gel is most commonly used for DNA separation.

Gel Electrophoresis Principles

DNA moves towards the cathode electrode.
Negative charge of DNA allows it to be separated by gel electrophoresis.
Buffer solution provides a medium for electrical conductivity.
Ethidium bromide is commonly used to visualize DNA in a gel after electrophoresis.
Loading dye makes DNA visible under UV light.

Gel Electrophoresis Types

Polyacrylamide gel provides higher resolution than agarose gel.

SDS-PAGE

SDS (sodium dodecyl sulfate) denatures proteins and gives them a uniform negative charge.

Gel Electrophoresis Resolution

Resolution refers to the ability to distinguish between two closely spaced DNA fragments.

Molecular Weight Markers

Molecular weight markers are used to determine the molecular weight of proteins.

RNA Separation

Agarose gel electrophoresis with formaldehyde is typically used for RNA separation.

Verifying Gel Electrophoresis Results

Accuracy can be verified by comparing with known standards.

Gel Electrophoresis Preparation

A comb is used to create wells for loading samples during gel preparation.

Medical Diagnostics

Gel electrophoresis is commonly used to detect genetic mutations in medical diagnostics.

Chromatography Types

Adsorption chromatography uses a column containing silica gel or alumina.
Two main components required for all chromatographic methods: stationary phase and mobile phase.

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Description

Test your knowledge of High Performance Liquid Chromatography (HPLC), HPLC detectors, and Gel Electrophoresis principles, including their applications and characteristics.