Large Volume Preparative Chromatography

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Questions and Answers

What is the typical capacity range of large volume preparative protocols?

100-500 mL

300-1700 mL (correct)

1000-3000 mL

5000-7000 mL

Why are certain protocols habitually chosen?

For experimental protocols

For analytical protocols

For small volume preparative protocols

For large volume preparative protocols (correct)

What is the lower limit of the typical capacity of large volume preparative protocols?

300 mL (correct)

200 mL

400 mL

100 mL

What is the upper limit of the typical capacity of large volume preparative protocols?

1700 mL Signup and view all the answers

What type of protocols are being discussed?

Preparative protocols Signup and view all the answers

What is the angle measured with respect to?

Axis of rotation Signup and view all the answers

What is the result of a shorter sedimentation pathlength?

Decreased centrifugal force Signup and view all the answers

What is the angle of a rotor mentioned in the text?

40° Signup and view all the answers

What is the relationship between the angle of the rotor and the sedimentation pathlength?

A larger angle results in a shorter sedimentation pathlength Signup and view all the answers

What is the characteristic of a rotor with a fixed angle?

Constant centrifugal force Signup and view all the answers

What is the purpose of the hole in the bottom of the centrifugal tube?

To allow for the reuse of the tube Signup and view all the answers

What is the direction of the gradient displacement?

From lower to higher density parts Signup and view all the answers

How are the fractions collected from the tube?

From the top of the tube to the bottom Signup and view all the answers

What happens to the particles of different densities during centrifugation?

They are separated based on their densities Signup and view all the answers

What is the result of using a more dense solution in the centrifugal tube?

The gradient is displaced upward Signup and view all the answers

Which type of viruses are related to human diseases?

Retroviruses Signup and view all the answers

What is an example of a human disease caused by a retrovirus?

HIV Signup and view all the answers

Which of the following diseases is NOT caused by a retrovirus?

Tuberculosis Signup and view all the answers

What is another name for a type of virus that is related to human diseases?

Retrovirus Signup and view all the answers

How many types of viruses are related to human diseases?

Several Signup and view all the answers

What is the duration of the process of suspending the pellet and mixing it with a concentrated salt solution?

28 hours Signup and view all the answers

What is the final density of the mixture after resuspending the pellet?

1.21 g/mL Signup and view all the answers

What is the amount of the pellet in grams?

100000 g Signup and view all the answers

What is the purpose of adding a concentrated salt solution to the pellet?

To increase the density of the mixture Signup and view all the answers

What is the resulting layer obtained after the process?

A floating layer of LDL Signup and view all the answers

Study Notes

Centrifugation Protocols

Large volume preparative protocols typically have capacities ranging from 300-1700 mL.
The sedimentation path length is shorter in a rotor with a fixed angle (e.g., 40°) compared to a vertical rotor.

Centrifugation Techniques

To collect fractions, a hole is punctured in the bottom of the centrifugation tube, and the fractions are collected from top to bottom, from higher to lower density parts of the gradient.
Displacement of the gradient upward is achieved using a more dense solution.

Characteristics of Retroviruses

Several retroviruses are related to human diseases, such as influenza and HIV.

Density Gradient Centrifugation

A floating layer of LDL is obtained, and the pellet is resuspended and adjusted to a final density of 1.21 g/mL.
The process involves centrifugation at 100,000 g for 28 hours, followed by resuspension and adjustment of the density to 1.063 g/mL.

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Description

This quiz covers the use of large volume preparative chromatography in protocols, typically with capacities ranging from 300-1700 mL. It's a crucial step in many laboratory processes.