UF MCB3020 EXAM 3 Bacusmo
Questions and Answers 100% Verified
Biotechnology - ANSWER - use of organisms to form useful products
Genetic engineering - ANSWER - The deliberate modification of an organisms genome
sequence
Recombinant DNA technology - ANSWER - procedures used to carry out genetic engineering
Cloning - ANSWER - generation of a large number of genetically identical DNA molecules
What are the 5 basic steps of cloning? - ANSWER - 1. Isolate DNA to be cloned
2. Use restriction enzymes to "chop up" the DNA into fragments
3. Insert the fragments into a cloning vector, creating recombinant DNA
4. Insert the recombinant DNA into a new host, such as E. coli
5. Culture host cell, growing many identical copies.
What are restriction enzymes? - ANSWER - A type of endonuclease that allows scientists to cut
up DNA at specific sites.
What is a cloning vector? - ANSWER - Provides means for transferring a gene of interest to a
host organism during the cloning process
What are the 3 things every "good" cloning vector must have? - ANSWER - •an origin of
replication
•a selectable marker
•a multicloning site (MCS)or polylinker
,True or False: Restriction enzymes sometimes make staggered cuts, which produce single-
stranded DNA sequences known as "sticky ends". - ANSWER - TRUE
What enzyme forms covalent bonds between the cloned gene and the plasmid, creating
recombinant DNA? - ANSWER - Ligase
What is reverse transcriptase? - ANSWER - When double-stranded DNA is created by a single-
stranded RNA.
What is the process or constructing cDNA? What is the goal of this process? - ANSWER - Goal:
make DNA from RNA template
1. Apply a short poly- T primer to RNA template
2. Add reverse transcriptase and the 4 nucleotides
3. Add RNaseH to cut up the RNA and regenerate the RNA primers.
4. Add DNA polymerase and DNA ligase to synthesize the new strand
What is gel electrophoresis? Why is it important? - ANSWER - A technique that separates
nucleic acids and proteins on the basis of their size and electrical charge.
It is important because it determines which fragments are larger than others and tells us the
approximate size for each fragment.
Nucleic acids have ____________ charge due to their _______________ _____________, so
they migrate through the electric field towards the _____________ electrode. - ANSWER -
negative; Phosphate backbones ; positive
True or False: Shorter DNA will travel farther in gel electrophoresis because smaller restriction
fragments move faster. - ANSWER - TRUE
What dye is used to stain DNA? - ANSWER - Ethidium bromide
What is PCR? - ANSWER - Polymerase Chain Reaction; A technique for quickly and easily
making many copies of even a very small amount of DNA.
, Where does Taq polymerase come from? - ANSWER - Thermophilic bacterium found in
hydrothermal vents (Thermus aquaticus)
What are the 4 things required in PCR? - ANSWER - •primers
•target DNA
•thermostable DNA polymerase (e.g. Taq polymerase)
*each of the four deoxyribonucleotide triphosphates
True or False: In PCR, hydrogen bonds of double-stranded DNA molecules are broken by
subjecting the DNA to high levels of radiation. - ANSWER - False; under high levels of HEAT
If you perform 6 cycles of PCR on a single double-stranded DNA molecule. How many copies
will you have? - ANSWER - 64 double-stranded copies (Each cycle the DNA content doubles)
Plasmids - ANSWER - self-replicating piece of extrachromosomal DNA
KNOW: this is the most commonly used & it's only in prokaryotes
Bacteriophages are - ANSWER - A virus that infects bacteria
Cosmids - ANSWER - A hybrid between a plasmid and a phage
artificial chromosomes - ANSWER - Synthetic chromosomes that contain fragments of DNA
integrated into a host chromosome
What is a genomic library? - ANSWER - a collection of the total genomic DNA from a single
organism
How are cloning vector DNA introduced into bacterial hosts? - ANSWER - Transformation: the
uptake of naked DNA from the environment
electroporation - ANSWER - (Related to Transformation) A technique that makes a cell
competent to pick up DNA from the environment by applying an electrical shock to it
Questions and Answers 100% Verified
Biotechnology - ANSWER - use of organisms to form useful products
Genetic engineering - ANSWER - The deliberate modification of an organisms genome
sequence
Recombinant DNA technology - ANSWER - procedures used to carry out genetic engineering
Cloning - ANSWER - generation of a large number of genetically identical DNA molecules
What are the 5 basic steps of cloning? - ANSWER - 1. Isolate DNA to be cloned
2. Use restriction enzymes to "chop up" the DNA into fragments
3. Insert the fragments into a cloning vector, creating recombinant DNA
4. Insert the recombinant DNA into a new host, such as E. coli
5. Culture host cell, growing many identical copies.
What are restriction enzymes? - ANSWER - A type of endonuclease that allows scientists to cut
up DNA at specific sites.
What is a cloning vector? - ANSWER - Provides means for transferring a gene of interest to a
host organism during the cloning process
What are the 3 things every "good" cloning vector must have? - ANSWER - •an origin of
replication
•a selectable marker
•a multicloning site (MCS)or polylinker
,True or False: Restriction enzymes sometimes make staggered cuts, which produce single-
stranded DNA sequences known as "sticky ends". - ANSWER - TRUE
What enzyme forms covalent bonds between the cloned gene and the plasmid, creating
recombinant DNA? - ANSWER - Ligase
What is reverse transcriptase? - ANSWER - When double-stranded DNA is created by a single-
stranded RNA.
What is the process or constructing cDNA? What is the goal of this process? - ANSWER - Goal:
make DNA from RNA template
1. Apply a short poly- T primer to RNA template
2. Add reverse transcriptase and the 4 nucleotides
3. Add RNaseH to cut up the RNA and regenerate the RNA primers.
4. Add DNA polymerase and DNA ligase to synthesize the new strand
What is gel electrophoresis? Why is it important? - ANSWER - A technique that separates
nucleic acids and proteins on the basis of their size and electrical charge.
It is important because it determines which fragments are larger than others and tells us the
approximate size for each fragment.
Nucleic acids have ____________ charge due to their _______________ _____________, so
they migrate through the electric field towards the _____________ electrode. - ANSWER -
negative; Phosphate backbones ; positive
True or False: Shorter DNA will travel farther in gel electrophoresis because smaller restriction
fragments move faster. - ANSWER - TRUE
What dye is used to stain DNA? - ANSWER - Ethidium bromide
What is PCR? - ANSWER - Polymerase Chain Reaction; A technique for quickly and easily
making many copies of even a very small amount of DNA.
, Where does Taq polymerase come from? - ANSWER - Thermophilic bacterium found in
hydrothermal vents (Thermus aquaticus)
What are the 4 things required in PCR? - ANSWER - •primers
•target DNA
•thermostable DNA polymerase (e.g. Taq polymerase)
*each of the four deoxyribonucleotide triphosphates
True or False: In PCR, hydrogen bonds of double-stranded DNA molecules are broken by
subjecting the DNA to high levels of radiation. - ANSWER - False; under high levels of HEAT
If you perform 6 cycles of PCR on a single double-stranded DNA molecule. How many copies
will you have? - ANSWER - 64 double-stranded copies (Each cycle the DNA content doubles)
Plasmids - ANSWER - self-replicating piece of extrachromosomal DNA
KNOW: this is the most commonly used & it's only in prokaryotes
Bacteriophages are - ANSWER - A virus that infects bacteria
Cosmids - ANSWER - A hybrid between a plasmid and a phage
artificial chromosomes - ANSWER - Synthetic chromosomes that contain fragments of DNA
integrated into a host chromosome
What is a genomic library? - ANSWER - a collection of the total genomic DNA from a single
organism
How are cloning vector DNA introduced into bacterial hosts? - ANSWER - Transformation: the
uptake of naked DNA from the environment
electroporation - ANSWER - (Related to Transformation) A technique that makes a cell
competent to pick up DNA from the environment by applying an electrical shock to it
