RNA Blotting Techniques Quiz

Podcast

Play an AI-generated podcast conversation about this lesson

Questions and Answers

What was the procedure devised by Alwine et al. (1978) for?

Determining the number of genes present on a given DNA

Transferring proteins from acrylamide gels to membranes

Transferring DNA bands from gel to chemically reactive paper

Transferring RNA bands from gel to chemically reactive paper (correct)

What is the theoretical advantage of Dot blotting?

Transferring RNA bands from gel to chemically reactive paper

Determining the number of genes through mRNA present on a given DNA (correct)

Modifying Southern and Northern blotting techniques

Transferring proteins from acrylamide gels to membranes

What are the drawbacks of Northern blotting?

Each gene may give rise to two or more RNA transcripts, the presence of introns

Each gene may give rise to two or more RNA transcripts, the absence of exons and introns

Each gene may give rise to two or more RNA transcripts, the presence of exons and introns (correct)

Each gene may give rise to only one RNA transcript, the presence of exons and introns

Which technique is used to transfer proteins from acrylamide gels to membranes?

Southern blotting Signup and view all the answers

Who coined the term 'western' blotting?

Burnette 1981 Signup and view all the answers

What technique is a modification of Southern and Northern blotting techniques?

Western blotting Signup and view all the answers

Who is the scientist that developed the Southern blotting technique in 1975?

Southern Signup and view all the answers

What is the first nucleic acid blotting procedure developed in 1975?

Southern blotting Signup and view all the answers

In Southern blotting, which type of DNA migrates towards the anode?

Genomic DNA Signup and view all the answers

What happens to smaller DNA fragments in Southern blotting?

They move towards the cathode Signup and view all the answers

After denaturation and transfer in Southern blotting, DNA fragments are transferred to which material?

Nitrocellulose or nylon paper Signup and view all the answers

What is the purpose of Southern blotting?

To analyze DNA fragments Signup and view all the answers

What happens to DNA when exposed to heat (80°C) in a Southern blotting technique?

It becomes negatively charged and moves faster on nitrocellulose, which is an exact replica. Signup and view all the answers

What helps draw the transfer buffer through the gel in Southern blotting?

The hybridization membrane immobilized on the absorbent material. Signup and view all the answers

What are the probes used in Southern blotting exposed to the nitrocellulose or nylon paper?

Labeled cDNA probes. Signup and view all the answers

What does the autoradiograph reveal in a Southern blotting technique?

Specific bands corresponding to the DNA fragments recognized by labeled cDNA probes. Signup and view all the answers

What is a variant of Southern blotting used for specific analysis?

Northern blotting used for RNA analysis. Signup and view all the answers

What is the purpose of hybridization membrane in Southern blotting?

To provide a surface for DNA probes to hybridize with complementary DNA molecules. Signup and view all the answers

Study Notes

Blotting Techniques

Western blotting is used to transfer proteins from acrylamide gels to membranes.
Northern blotting is a technique for determining the number of genes (through mRNA) present on a given DNA.
Southern blotting is a procedure in which RNA bands are blot transferred from the gel onto chemically reactive paper, where they are bound covalently.
Dot blotting is a modification of Southern and Northern blotting techniques.

Southern Blotting Procedure

Genomic DNA is isolated from cells/tissues and digested with one or more restriction enzymes.
The mixture is loaded into a well in an agarose or polyacrylamide gel and then subjected to electrophoresis.
DNA, being negatively charged, migrates towards the anode; smaller DNA fragments move faster.
The separated DNA molecules are denatured by exposure to a mild alkali and transferred to nitrocellulose or nylon paper.

Transfer of DNA Molecules

A nitrocellulose or nylon paper is sandwiched between the gel and a stack of paper towels, which draws the transfer buffer through the gel by capillary action.
The DNA molecules are carried out of the gel by the buffer flow and immobilized on the membrane.

Hybridization and Autoradiography

The nitrocellulose or nylon paper is exposed to labeled cDNA probes, which hybridize with complementary DNA molecules on the paper.
The paper, after thorough washing, is exposed to X-ray film to develop autoradiograph, revealing specific bands corresponding to the DNA fragments recognized by the probe.

Variants of Blotting Techniques

Eastern blotting is a variant of Southern blotting used for specific analysis.
Dot blotting is a variant of Southern and Northern blotting techniques.

Studying That Suits You

Use AI to generate personalized quizzes and flashcards to suit your learning preferences.

Description

Test your knowledge on RNA blotting techniques such as Northern blotting, dot blotting, and negative blotting with this quiz. Learn about the procedure of transferring RNA bands from a gel onto chemically reactive paper.