Biotechnology - Answer-Use of living organisms to create products that are useful to humans
Genetic Engineering - Answer-Deliberate modification of an organisms genome sequence
Recombinant DNA Technology - Answer-Set of procedures used to combine genetic material from multiple sources and cre...
MCB 3020 Exam 3 Study Guide
Questions and Answers
Biotechnology - Answer-Use of living organisms to create products that are useful to
humans
Genetic Engineering - Answer-Deliberate modification of an organisms genome
sequence
Recombinant DNA Technology - Answer-Set of procedures used to combine genetic
material from multiple sources and create DNA sequences that are otherwise not found
in biological organisms
Cloning - Answer-Generation of a larger number of identical DNA molecules from a
single DNA molecule
5 Basic Steps of Cloning - Answer-1. Isolate DNA to be cloned
2. Use restriction enzymes to "chop up" DNA into fragments
3. Insert fragments into cloning vector, creating recombinant DNA
4. Insert recombinant DNA into new host, such as E. coli
5. Culture host cell, growing many identical copies
Restriction Enzymes - Answer-Type of endonuclease that allows scientists to cut up
DNA at specific sites
T/F: Restriction enzymes sometimes make staggered cuts, which produce single-
stranded DNA sequences known as "sticky ends" - Answer-True
What enzyme forms covalent bonds between cloned gene and plasmid, creating
recombinant DNA? - Answer-Ligase
What is reverse transcriptase? - Answer-When double-stranded DNA is created from
single-stranded RNA
What is the process of constructing cDNA? What is the goal of this process? - Answer-
Goal: make DNA from RNA template
1. Apply short poly-T primer to RNA template
2. Add reverse transcriptase and 4 nucleotides
3. Add RNaseH to cut up RNA and regenerate RNA primers
4. Add DNA pol and DNA ligase to synthesize new strand
What is gel electrophoresis? Why is it important? - Answer-A technique that separates
nucleic acids and proteins based on the basis of their size and electrical charge
, Important because it determines which fragments are larger than others and tells us the
approximate size of each fragment
Nucleic acids have ___ charge to due their ____ ____, so they migrate through electric
field towards ____ electrode - Answer-Negative; phosphate backbones; positive
T/F: 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-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-DNA of interest, primer, DNA
polymerase, deoxyribonucleotide triphosphates
T/F: In PCR, hydrogen bonds of double-stranded DNA molecules are broken by
subjecting DNA to high levels of radiation - Answer-False; high levels of HEAT
If you perform 6 cycles of PCR on a double-stranded DNA molecule how many copies
will you have? - Answer-64; each cycle it doubles
What is a cloning vector? - Answer-Provides means for transferring a gene of interest to
a host organism during cloning process
What are the 3 things every "good" cloning vector must have? - Answer-An origin of
replication, selectable marker, unique restriction sites calls POLYLINKERS or MCSs
Describe the following types of cloning vectors - Answer-- Plasmids: self-replicating
piece of extrachromosomal DNA (most commonly used and its only in prokaryotes)
- Bacteriophages: virus that infects bacteria
- Cosmids: hybrid between plasmid and phage
- Artificial chromosomes: synthetic chromosomes that contain fragments of DNA
integrated into host chromosome
What is a genomic library? - Answer-Collection of an organism's entire genomic DNA
How are cloning vector DNA introduced into bacterial hosts? - Answer-Transformation:
uptake of naked DNA from environment
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