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UF MCB3020 EXAM 3 Bacusmo Questions and Answers 100% VerifiedUF MCB3020 EXAM 3 Bacusmo Questions and Answers 100% VerifiedUF MCB3020 EXAM 3 Bacusmo Questions and Answers 100% VerifiedUF MCB3020 EXAM 3 Bacusmo Questions and Answers 100% Verified Biotechnology - ANSWER - use of organisms to form u...

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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

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