MBIOS 303: chapter 20 (exam 2)
In dideoxyribonucleotides, _______ oxygens are removed from the sugar compared with ribose.
2
Which component in the CRISPR-Cas9 system makes a double-strand break in DNA?
Cas9
Polymerase chain reaction was developed by ______.
Kary Mullis
An early method of DNA sequencing that involved the base-specific chemical cleavage of DNA was developed by ______.
Maxam & Glibert
This technique is used to identify a specific RNA molecule within a mixture of RNA molecules.
Northern blotting
In PCR, the template DNA is ______.
The DNA to be amplified
True or false: Amplifying a gene by PCR results in many copies, similar to cloning using a vector and host cell.
True
True or false: Chromosomal DNA is a common source of cloned DNA.
True
In PCR, each cycle uses the products of the previous cycle as templates. What do you call this?
a chain reaction
Methods that produce two or more genetically identical individuals are referred to as reproductive _______
cloning
Cells that can take up DNA from the medium are known as _______ cells
competent
In PCR, primer extension refers to the synthesis of ______ starting at the primers.
complementary DNA
During the initial phase of a real-time PCR experiment, called the _______ phase, the amount of PCR products is small and reagents are not limiting, so the amount of product nearly doubles with each cycle.
exponential
When using PCR to amplify DNA, short oligonucleotides called primers ______.
flank the region of DNA to be amplified
Knowing the sequence of the DNA ______ the gene of interest allows scientists to design appropriate primers.
flanking
Which process will result in a point mutation in the target gene when using the CRISPR-Cas technology for gene editing?
homologous recombination repair
A cell that harbors a vector is called a(n) _______ _______
host cell
The natural function of the CRISPR-Cas system in bacteria is to ______.
provide defense against bacteriophages
The technique that allows one to assess the amount of DNA produced during a PCR amplification as it is happening is called _______-_______ PCR.
real-time
A vector that contains a piece of chromosomal DNA is referred to as a(n) _______ vector.
recombinant
Stem cells that can give rise to all the cell types in the adult organism are said to be
totipotent
When the CRISPR-Cas system is used for gene mutagenesis, which two components are combined in the sgRNA?
tracrRNA crRNA
Which technique is used to identify a particular protein in a mixture of proteins?
western blotting
You wish to determine if a protein is made at a particular stage of development. Which technique would you use?
western blotting
Order the following steps in cloning a gene, placing the first step at the top. Instructions Choice 1 of 3. Ligation by DNA ligase produces recombinant vectors. Choice 2 of 3. Chromosomal DNA is isolated and cut with a restriction enzyme; the plasmid DNA is cut with the same enzyme. Choice 3 of 3. The digested chromosomal DNA and plasmid DNA are incubated together.
1. Chromosomal DNA is isolated and cut with a restriction enzyme; the plasmid DNA is cut with the same enzyme. 2. The digested chromosomal DNA and plasmid DNA are incubated together. 3. Ligation by DNA ligase produces recombinant vectors.
Order the steps in one cycle of a PCR reaction, placing the first step at the top. - primer extension - denaturation - primer annealing
1. denaturation 2. primer annealing 3. primer extension
In CRISPR-Cas9 mutagenesis, which type of change is observed in the target gene when nonhomologous end joining (NHEJ) repairs the double-strand break?
A small deletion causes a frameshift mutation.
In PCR, the two primers bind to specific sites in the _______ and the flank the gene to be amplified.
DNA
This technique enables researchers to determine the DNA bases in genes and other chromosomal regions.
DNA sequencing
A significant breakthrough in research in the use of stem cells for therapy was achieved in 1998 when two separate teams showed that ______.
ES and EG cells can be propagated in the lab
How is ethidium bromide used to analyze amplification by PCR?
Ethidium bromide stains DNA, allowing the size of a fragment in a gel to be determined.
Match the phase of real-time PCR with the accumulation of products.
Exponential- The amount of product nearly doubles with each cycle but may be difficult to detect as the amounts are small. Linear- Accumulation of the product shows a directly proportional relationship to cycle number. Plateau- Accumulation of the product levels off as one or more reagents are used up.
True or false: Reproductive cloning can only occur as a result of laboratory manipulations.
False
How is site-directed mutagenesis useful in the study of genes and proteins?
The mutated gene can be introduced into a living organism to see how the mutation affects the organism.
When cloning a gene, why must the chromosomal DNA and the plasmid DNA be cut with the same restriction enzyme?
The sticky ends of the plasmid DNA will be complementary to the sticky ends of the chromosomal DNA.
In PCR, why do the primers bind to specific sites in the DNA on either side of the gene of interest?
They are complementary to the flanking sequences.
Which of the following are common characteristics of stem cells?
They can differentiate into one or more specialized cell types. They have the capacity to divide.
In gene cloning, what is the vector?
a small DNA molecule that can replicate independently within a host cell
What is a plasmid?
a small circular DNA molecule often used as a vector in gene cloning
What type of apparatus does one need to quantify the DNA produced during real-time PCR?
a thermocycler that can detect fluorescence
A totipotent cell can give rise to ______ of the cell types in an adult organism.
all
Site-directed mutagenesis allow a researcher to make a mutation ______.
at a specific sequence of DNA
DNA sequencing allows researchers to determine the _______ _______ of DNA found in genes and other chromosomal regions.
base order (nucleotide sequence)
Stem cells could be used to treat conditions of the _______ , such as Parkinson's disease.
brain
How is the amount of DNA produced during real-time PCR measured?
by measuring the fluorescence emitted by the probe added to the PCR mixture
A DNA library made with DNA generated by reverse transcriptase is called a(n) _______ library.
cDNA
If you wanted a collection of only transcribed DNA, you would construct a ______.
cDNA libary
Using stem cells to repair joints damaged by injury or arthritis would be useful in which tissue type?
cartilage
A particular gene to be cloned is often isolated from ______.
chromosomal DNA
To make many copies of a gene, you would _______ that gene.
clone
If a gene is amplified by PCR so that there are many copies, it can be said to be ______.
cloned
A recombinant vector ______.
contains a piece of chromosomal DNA
A vector requires an origin of replication so that it can be ______.
copied many times by the host cell
Restriction endonucleases are used in gene cloning to ______.
cut the DNA backbone prior to inserting the DNA to be cloned
Reverse transcriptase PCR can be used to ______.
detect and quantify the amount of a specific RNA
Scientists are mainly interested in stem cells to help them understand basic genetic mechanisms that underlie the process of _______, and for their potential to treat human _______.
development; disease
If the oxygens on carbons 2 and 3 of the sugar of a nucleotide have been removed, as shown in the image, the nucleotide is referred to as a(n)
dideoxyribonucleotide (ddNTP)
When CRISPR-Cas is used for gene mutagenesis, what type of change does Cas9 make in the gene of interest?
double-strand break
A technique for identifying proteins that bind to DNA or RNA is called the _______ _______ _______ assay (EMSA).
electrophoretic mobility shift
The early mammalian embryo contains _______ _______ cells, which are pluripotent cells found in the inner cell mass of the blastocyst.
embryonic stem
Which fluoresces under UV light and can be used to detect DNA in a gel?
ethidium bromide
What is the term that describes a cell that contains a DNA cloning vector?
host cell
Primers are chosen for PCR based on ______.
knowing the DNA sequences flanking the gene of interest
In PCR, the temperature must be ______ from the denaturation temperature in order for primers to anneal.
lowered
In gene cloning, cells are treated with agents that ______, creating competent cells.
make them permeable to DNA
Hematopoietic stem cells populate blood and lymphoid tissues and are ______.
multipotent
Stem cells that can differentiate into several cell types, but far fewer than an embryonic stem cell, are said to be _______
multipotent
In gene mutagenesis by CRISPR-Cas, a DNA break can be repaired by _______ end _______ or by homologous_______ _______ .
nonhomologous; joining; recombination repair
This technique is used to identify a specific RNA molecule within a mixture of RNA molecules.
northern blotting
What do you call the DNA sequence in a vector that allows the replication enzymes of the host cell to make multiple copies of the vector?
origin of replication
A small DNA molecule that can replicate independently within a host cell and thus make many copies of an inserted gene is called a(n) _______
plasmid
A small circular DNA molecule that is often used as a vector in gene cloning is called a(n) _______
plasmid
Embryonic stem cells are _______ , which means that they can differentiate into almost every cell type of the body, but a single embryonic stem cell has lost the ability to produce an entire, intact individu
pluripotent
stem cell that can differentiate into almost every cell type in the body, but has lost the ability to produce an entire, intact individual is called ______.
pluripotent
The step in PCR that occurs when Taq polymerase starts at the primers and catalyzes the synthesis of complementary DNA is called _______ _______
primer extension
Short oligonucleotides that flank the region of DNA to be amplified by PCR are called _______
primera
What is the technique that allows one to determine the amount of template DNA present when the PCR cycles began?
real-time PCR
The technology using in vitro molecular techniques that combine DNA fragments to produce novel arrangements is called _______ DNA technology.
recombinant
Enzymes that bind to a specific DNA sequence and cut the DNA backbone are called _______ _______
restriction enzymes
You have a piece of RNA, and you want to synthesize a complementary strand of DNA. Which enzyme would you use?
reverse transcriptas
The enzyme _______ _______ is used when PCR is employed to detect and quantify the amount of a specific RNA.
reverse transcriptase
The enzyme that uses RNA as a template to make a complementary strand of DNA is called _______ _______
reverse transcriptase
Primer annealing occurs when ______.
short oligonucleotides bind to DNA flanking the gene of interest
When the CRISPR-Cas system is used for gene mutagenesis, tracrRNA and crRNA are linked together in a molecule called the _______ _______ RNA (sgRNA)
single guide
If a scientist wanted to determine how a specific mutation in a gene of interest affected an organism, which technique would be most useful?
site-directed mutagenesis
The technique that allows a researcher to produce a mutation at a specific sequence is called
site-directed mutagenesis
Prior to the cloning of Dolly, it was believed that the chromosomes of _______ cells of mammals had incurred irreversible genetic changes that rendered them unsuitable for cloning. However, Dolly was cloned using DNA from _______ cells.
somatic; somatic
In the CRISPR-Cas system, what part of the sgRNA is designed to be complementary to the gene to be mutated?
spacer region
The Maxam and Gilbert method of DNA sequencing used chemicals that cleaved the DNA at ______.
specific bases
The cells that construct our bodies from a fertilized egg are called _______ cells.
stem
Which component of bone marrow is useful to patients with certain types of cancers?
stem cells
With the exception of those in the blood, other types of adult _______ _______ are difficult to locate and to remove in sufficient numbers for transplantation.
stem cells
In PCR, the DNA to be amplified is called the _______ DNA.
template
Embryonic stem cells are found in ______.
the blastocyst
Which technique is useful for studying protein-DNA interactions?
the electrophoretic mobility shift assay
What is recombinant DNA technology?
the production of new arrangements of DNA
To perform PCR, a machine called a(n) _______ automates the timing of each cycle.
thermocycler
To perform many cycles of PCR, you need a machine that can change temperatures at exact times. What do you call this machine?
thermocycler
What is the purpose of Northern blotting?
to identify a specific RNA molecule within a mixture of RNA molecules
What is the purpose of gene cloning?
to produce many copies of a DNA molecule of interest
