Molecular Cloning

Question 1

Refers to isolating and copying a particular DNA segment of interest for further study

Answer 1

Molecular Cloning

Question 2

A laboratory process of making millions of copies of a single molecule, most often DNA.

Answer 2

Molecular Cloning

Question 3

Molecular cloning is used in what.

Answer 3

To amplify or make many copies of DNA and to perform genetic tests on small samples of blood, saliva, or other tissues from patients

Question 3

It’s different from cellular cloning or organism cloning that are used in reproductive genetics

Answer 3

Animal Cloning

Question 4

What are the two senses wherein DNA cloning is used.

Answer 4

It refers to the act of making many identical copies of a DNA molecule and the isolation of a particular stretch of DNA (often a particular gene) from the rest of a cell’s DNA.

Question 5

Give 6 types of Gene cloning.

Answer 5

1. Starting DNA/RNA material – extraction/purification
2. Polymerase chain reaction
3. Restriction enzymes
4. Cloning vectors
5. Bacterial transformation
6. DNA sequencing

Question 6

A basic method used in molecular biology

Answer 6

Extraction of DNA and RNA

Question 7

Is important for a wide range of research and clinical applications.

Answer 7

High-quality, highly pure nucleic acid

Question 7

What can be extracted and purified from bacterial and mammalian cells.

Answer 7

Genomic DNA, Plasmid DNA, and Total RNA

Question 7

An initial step in many molecular biology and genomic workflows

Answer 7

Nucleic acid purification

Question 8

Give examples of sources from which Genomic DNA, plasmid DNA, and total RNA can be extracted.

Answer 8

• bacterial and mammalian cells
• blood
• mammalian tissue
• fungal tissue
• plant tissue
• plasma
• serum

Question 8

Isolation of nucleic acid involves

Answer 8

Lysis of cell membranes or sample homogenization, followed by the removal of proteins, enzymes, detergents, salts, and lipids

Question 9

Difference between chromosomal DNA and plasmid DNA

Answer 9

Plasmid DNA only contains extra genes that are not really necessary for the organism existence, but chromosomal DNA carries all of the information needed for the growth of the organism, development of the organism, and reproduction.

Question 10

How does nucleic acid extraction and isolation starts

Answer 10

With disrupting the cellular structures containing the genetic material. This is done by adding a compatible detergent, mechanical disruption, and/or heat.

Question 11

Approaches in DNA/RNA Extraction and Purification

Answer 11

• Alkaline extraction
• Phenol-chloroform extraction
• Cesium chloride (CsCl) density gradient centrifugation
• Oligo(dT)-cellulose chromatography
• Silica matrice
• Glass beads

Question 11

Nucleic acid isolation by organic extraction involves

Answer 11

Addition of phenol and guanidine isothiocyanate to separate the DNA, RNA, and proteins into different organic phases

Question 12

Samples are lysed with a phenol-chloroform solution and centrifuged.

Answer 12

Phenol-chloroform extraction

Question 13

Three phases that can be seen after centrifugation

Answer 13

An upper aqueous phase containing nucleic acids, a lower organic phase of lipids, and an interphase comprising proteins