Chapter 20: Molecular Technologies
plasmid
A small circular DNA molecule that is often used as a vector in gene cloning is called a(n)___________.
copied many times by the host cell
A vector requires an origin of replication so that it can be ______.
a million
After 20 PCR cycles, a DNA sequence may be amplified ______-fold.
about 20
After ______ cycles of PCR, a DNA sequence may be amplified a million-fold.
human cell lines adult mice mouse embryos roundworms plant cells
CRISPR-Cas technology has been used to mutate genes in ______.
Which component in the CRISPR-Cas9 system makes a double-strand break in DNA?
Cas9
DNA nucleotides
DNA sequencing enables researchers to determine the order of ______ ______ in a gene.
The mutated gene can be introduced into a living organism to see how the mutation affects the organism.
How is site-directed mutagenesis useful in the study of genes and proteins?
Site-directed mutagenesis
If a scientist wanted to determine how a specific mutation in a gene of interest affected an organism, what technique would be most useful?
polymerase chain reaction
In 1985, Kary Mullis developed a way to copy DNA without vectors or host cells. This technique is called_________ ____________ ____________.
A chain reaction
In PCR, each cycle uses the products of the previous cycle as templates. What do you call this?
template
In PCR, the DNA to be amplified is called the ____________ DNA.
the DNA to be amplified
In PCR, the template DNA is ______.
DNA
In PCR, the two primers bind to specific sites in the_______ and flank the gene to be amplified.
They are complementary to the flanking sequences.
In PCR, why do the primers bind to specific sites in the DNA on either side of the gene of interest?
fluorescent dye
In automated sequencing, each dideoxyribonucleotide is labeled with a different colored ______.
A small DNA molecule that can replicate independently within a host cell
In gene cloning, what is the vector?
1. Many copies of a single-stranded vector containing the gene of interest are acquired 2. Primers are added. 3. High concentrations of all four unlabeled deoxyribonucleotides are added. 4. Low concentrations of all four fluorescently labeled dideoxyribonucleotides and DNA polymerase are added. 5. Electrophoresis separates DNA strands according to their lengths. 6. A laser and fluorescence detector are used to determine the color associated with each DNA strand.
Order the steps in DNA sequencing, putting the first step at the top.
1. Exponential 2. Linear 3. Plateau
Real-time PCR goes through three main phases with regard to product accumulation. Place them in order from first to last, with the first step at the top.
mRNA dNTPs Poly-dT primer Reverse transcriptase
Select the reagents needed to make cDNA.
primers
Short oligonucleotides that flank the region of DNA to be amplified by PCR are called _______.
at a specific sequence of DNA
Site-directed mutagenesis allow a researcher to make a mutation ______.
reverse transcriptase
The enzyme that uses RNA as a template to make a complementary strand of DNA is called____________ _________.
to provide defense against bacteriophages
The natural function of the CRISPR-Cas system in bacteria is to ______.
sequence of DNA
The use of dideoxyribonucleotides with different colored fluorescent dyes allows the detection of the ______.
A. High B. Medium C. Low
This figure shows a real-time PCR experiment carried out at low, medium, and high concentrations of starting template DNA. Please match each curve (designated by a letter) with the corresponding amount of starting template. Fluorescence values are plotted on the Y axis.
DNA sequencing
This technique enables researchers to determine the DNA bases in genes and other chromosomal regions.
Northern blotting
This technique is used to identify a specific RNA molecule within a mixture of RNA molecules.
high concentrations of unlabeled deoxyribonucleotides and low concentrations of labeled dideoxyribonucleotides
To ensure that the automated sequencing is accurate, it is important that the sequencing mixture contains ______.
Thermocycler
To perform many cycles of PCR, you need a machine that can change temperatures at exact times. What do you call this machine?
True
True or false: Chromosomal DNA is a common source of cloned DNA.
Origin of replication
What do you call the DNA sequence in a vector that allows the replication enzymes of the cell to make lots of copies of the vector?
A collection of recombinant vectors
What is a DNA library?
A molecule that carries the DNA to be cloned
What is a cloning vector?
A cDNA molecule contains the coding regions but not introns.
What is an advantage of using cDNA in cloning?
The production of new arrangements of DNA
What is recombinant DNA technology?
To identify a specific RNA molecule within a mixture of RNA molecules
What is the purpose of Northern blotting?
To produce many copies of a DNA molecule of interest
What is the purpose of gene cloning?
Real-time PCR
What is the technique that allows one to determine the amount of template DNA present when the PCR cycles began?
Host cell
What is the term that describes a cell that contains a DNA cloning vector?
Western blotting
What technique is used to identify a particular protein in a mixture of proteins?
flank the region of DNA to be amplified
When using PCR to amplify DNA, short oligonucleotides called primers ______.
Cloned genes can be used in trials of gene therapy. Cloned genes can be introduced into bacteria to make medicines. The expression of a cloned gene can be used to discover its cellular function.
Which of the following are common uses of gene cloning?
Cloned genes can be used in trials of gene therapy. The expression of a cloned gene can be used to discover its cellular function. Cloned genes can be introduced into bacteria to make medicines.
Which of the following are common uses of gene cloning?
The expression of a cloned gene can be used to discover its cellular function. Cloned genes can be introduced into bacteria to make medicines. Cloned genes can be used in trials of gene therapy.
Which of the following are common uses of gene cloning?
Insulin
Which of the following is pharmaceutical product that is produced by bacteria expressing the human gene?
A cDNA molecule does not have introns, which can be quite large.
Why is cloning a cDNA molecule easier than cloning an entire eukaryotic gene?
It would be complementary to the poly-A tail at the 3' end of the mRNA.
Why would you use a poly-dT primer when making cDNA?
Reverse transcriptase
You have a piece of RNA, and you want to synthesize a complementary strand of DNA. What enzyme would you use?
clone
You would____________ a gene to make many copies of that gene.
site-directed mutagenesis
_________-_________ ___________allows a researcher to produce a mutation at a specific sequence.
real time
___________-_____________ PCR allows one to assess the amount of DNA produced during a PCR amplification as it is happening.
Recombinant
_______________ DNA technology uses in vitro molecular techniques that combine DNA fragments to produce novel arrangements.
1. Denaturation 2. Primer Annealing 3. Primer Extension
rder the steps in one cycle of a PCR reaction, putting the first step at the top.
cloning vector
A DNA molecule that acts as a carrier of DNA that is to be cloned is called a(n) ______.
host cell
A cell that harbors a vector is called a________ __________.
host cell
A cell that harbors a vector is called a__________ __________.
living
A major advantage of CRISPR-Cas technology over site-directed mutagenesis is that it can be used directly on_____________ cells.
chromosomal DNA
A particular gene to be cloned is often isolated from ______.
DNA library
A researcher may use restriction enzymes to digest the DNA of an organism. The fragments of DNA are then ligated individually into many vectors. This collection of recombinant vectors is called a__________ _____________.
vector
A small DNA molecule that can replicate independently within a host cell and thus make many copies of an inserted gene is called a_________.