MOL BIO LAB L9 (Semis- Blotting Techniques)

Question 1

Identify the first step in each blotting technique:
1) Southern blotting
2) Northern blotting
3) Western blotting

Answer 1

1) Southern blotting
- DNA sample is first digested with restriction enzymes

While in both:
2) Northern blotting
3) Western blotting
- RNA sample (In northern) or protein sample (in western) is first separated by gel electrophoresis.

Question 2

In Southern blotting, these cut the DNA at specific sequences

Answer 2

Restriction enzymes

Question 3

Blotting techniques are based on the principle of?

Answer 3

Hybridization

Question 4

Used to detect specific DNA sequences in a biological sample

Answer 4

Southern Blotting

Question 5

Used to detect specific proteins in a biological sample

Answer 5

Western Blotting

Question 6

Applications of Southern blotting

Answer 6

• Diagnose genetic diseases, such as cystic fibrosis and sickle cell anemia
• Identify transgenic organisms
• Detect DNA viruses.

Question 7

Refers to the specific binding of two complementary nucleic acid sequences or two complementary protein sequences

Answer 7

Hybridization

Question 8

In western blotting, the probe can be visualized using a variety of methods, such as?

Answer 8

chemiluminescence or fluorescence

Question 9

The three main types of blotting techniques

Answer 9

1) Southern blotting
2) Northern blotting
3) Western blotting

Question 10

Applications of Northern blotting

Answer 10

• Study gene expression
• Identify differentially expressed genes in different tissues or cell types
• Detect RNA viruses

Question 11

A set of laboratory methods used to detect and analyze specific molecules

Answer 11

Blotting Techniques

Question 12

Blotting techniques are widely used in?

Answer 12

molecular biology and biochemistry research

Question 13

ARRANGE IN ORDER: 1-6

SOUTHERN BLOTTING
__ The probe is visualized using a variety of methods such as autoradiography, or chemiluminescence

__ DNA fragments are transferred to a membrane

__ DNA samples are first digested with restriction enzymes

__ If the target DNA sequence is present in the sample, the probe will bind to it

__ The membrane is probed with a labeled DNA probe

__ DNA fragments are separated by gel electrophoresis

Answer 13

6) The probe is visualized using a variety of methods such as autoradiography, or chemiluminescence

3) DNA fragments are transferred to a membrane

1) DNA samples are first digested with restriction enzymes

5) If the target DNA sequence is present in the sample, the probe will bind to it

4) The membrane is probed with a labeled DNA probe

2) DNA fragments are separated by gel electrophoresis

Question 14

In Southern blotting, DNA fragments are transferred to a membrane, such as?

Answer 14

nitrocellulose or nylon

Question 15

If the target protein is present in the sample, the _______ will bind to it. What blotting technique is this?

Answer 15

antibody; Western blotting

Question 16

This is used to probe the membrane and is specific to the target protein

Answer 16

labeled antibody

Question 17

Used to detect specific RNA sequences in a biological sample

Answer 17

Northern Blotting

Question 18

Applications of Western blotting

Answer 18

• Diagnose infectious diseases, such as HIV/AIDS and hepatitis C.
• Identify cancer biomarkers
• Study protein-protein interactions

Question 19

This is complementary to the target RNA sequence and is used to probe the membrane

Answer 19

labeled RNA probe

Question 20

In southern and northern blotting, the probe can be visualized using a variety of methods, such as?

Answer 20

Autoradiography or chemiluminescence

Question 21

In blotting techniques, DNA fragments or RNA/Protein samples are separated by?

Answer 21

Gel electrophoresis

Question 22

This is complementary to the target DNA sequence and is used to probe the membrane

Answer 22

labeled DNA probe