Chapter 20 - part of exam 2

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.

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

DNA sequencing allows researchers to determine the ________ of DNA found in genes and other chromosomal regions

Base sequence

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

Using stem cells to repair joints damaged by injury or arthritis would be useful in which tissue type?

Cartilage

Which component in the CRISPR-Cas9 system makes a double-strand break in DNA?

Cas9

To make many copies of a gene, you would ______ that gene.

Clone

Methods that produce two or more genetically identical individuals are referred to as reproductive

Cloning

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

Cells that can take up DNA from the medium are known as

Competent cells

In PCR, the two primers bind to specific sites in the ______ and flank the gene to be amplified.

DNA

This technique enables researchers to determine the DNA bases in genes and other chromosomal regions.

DNA sequencing

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 or ddNTPs

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.

A technique for identifying proteins that bind to DNA or RNA is called the _____________ assay (EMSA).

Electrophoretic mobility shift assay

The early mammalian embryo contains _______ cells, which are pluripotent cells found in the inner cell mass of the blastocyst.

Embryonic stem cells

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.

True or false: Reproductive cloning can only occur as a result of laboratory manipulations

False

Polymerase chain reaction was developed by ______.

Kary Mullis

In gene mutagenesis by CRISPR-Cas, a DNA break can be repaired by _______ end _______ or by homologous ______

Nonhomologous end joining Recombination repair

This technique is used to identify a specific RNA molecule within a mixture of RNA molecules.

Northern blotting

The step in PCR that occurs when Taq polymerase starts at the primers and catalyzes the synthesis of complementary DNA is called

Primer extension

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

The enzyme _____________ is used when PCR is employed to detect and quantify the amount of a specific RNA.

Reverse transciptase

When the CRISPR-Cas system is used for gene mutagenesis, tracrRNA and crRNA are linked together in a molecule called the

Single guide RNA (sgRNA)

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

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

Stem cells

Which component of bone marrow is useful to patients with certain types of cancers?

Stem cells

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.

common characteristics of stem cells

They have the capacity to divide. They can differentiate into one or more specialized cell types.

What is the purpose of Northern blotting?

To identify a specific RNA molecule within a mixture of RNA molecules

True or false: Chromosomal DNA is a common source of cloned DNA.

True

In dideoxyribonucleotides, ________ oxygens are removed from the sugar compared with ribose.

Two

Which technique is used to identify a particular protein in a mixture of proteins?

Western blotting

What is a plasmid?

a small circular DNA molecule often used as a vector in gene cloning

Embryonic stem cells are found in ______.

blastocyst

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 library

In PCR, each cycle uses the products of the previous cycle as templates. What do you call this?

chain reaction

A particular gene to be cloned is often isolated from ______.

chromosomal DNA

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.

Which fluoresces under UV light and can be used to detect DNA in a gel?

ethidium bromide

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

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

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 sequence 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

Stem cells that can differentiate into several cell types, but far fewer than an embryonic stem cell, are said to be

multipotent

A 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

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 individual.

pluripotent

Short oligonucleotides that flank the region of DNA to be amplified by PCR are called

primers

Enzymes that bind to a specific DNA sequence and cut the DNA backbone are called

restriction enzymes

The enzyme that uses RNA as a template to make a complementary strand of DNA is called

reverse transcriptase

You have a piece of RNA, and you want to synthesize a complementary strand of DNA. Which enzyme would you use?

reverse transcriptase

Primer annealing occurs when ______.

short oligonucleotides bind to complementary DNA flanking the gene of interest

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

In the CRISPR-Cas system, what part of the sgRNA is designed to be complementary to the gene to be mutated?

spacer region

Site-directed mutagenesis allow a researcher to make a mutation ______.

specific sequence of DNA

In PCR, the DNA to be amplified is called the ______ DNA.

template

Which technique is useful for studying protein-DNA interactions?

the electrophoretic mobility shift assay

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 gene cloning?

to produce many copies of a DNA molecule of interest

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 and crRNA

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)

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(n) ________.

vector or plasmid

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.

1. Chromosomal DNA is isolated and cut with a restriction enzyme; the plasmid 2. The digested chromosomal DNA and plasmid DNA are incubated together. 3. Ligation by DNA ligand produces recombinant vectors.

Order the steps that were used to create Dolly, the sheep cloned from somatic cell DNA, placing the first step at the top.

1. Mammary cells from an adult sheep were removed and grown in the lab; researchers extracted the nucleus from an egg cell from a different sheep. 2. Electrical pulses were used to fuse the diploid mammary cell with the enucleated egg cell. 3. The zygote began embryonic development. 4. The resulting embryo was implanted into the uterus of a surrogate mother sheep.

Order the steps in one cycle of a PCR reaction, placing the first step at the top.

1.denaturation 2.primer annealing 3.primer extension

In gene cloning, what is the vector?

A small DNA molecule that can replicate independently within a host cell