Genetics Lesson 20

short oligonucleotides bind to complementary DNA flanking the gene of interest

Primer annealing occurs when?

complementary

When DNA is made using RNA as the *starting* *material*, the DNA is called _________ DNA.

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

clone

You would _______ a gene to make many copies of that gene?

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 arrangement.

cDNA

A DNA library made with DNA *generated* by *reverse* *transcriptase* is called a ________ library.

cloning vector

A DNA molecule that *acts* *as* *a* *carrier* of *DNA* *that* *is* *to* *be* *cloned* is called a?

host cell

A cell that harbors a vector is called a _______ ________.

the DNA to be amplified

In PCR, the template DNA is?

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?

it covalently links the sugar-phosphate backbone of DNA

What does the enzyme DNA ligase do?

at least two different sources

A recombinant DNA molecule has covalently linked DNA fragments from?

primer extension

During PCR, the process of _____ _____ results when the Taq polymerase catalyzes the synthesis of complimentary DNA, starting at the primers.

reverse transcriptase

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

Recombinant DNA

A molecule that has covalently linked DNA fragments from at least two sources is called a _______ ___ molecule.

chromosomal DNA

A particular gene to be cloned is often isolated from?

selectable marker

A resistance gene that allows a host cell containing a vector to grow on a toxic substance is called a(n)?

oligonucleotide

A short strand of DNA, called a(n) ________, might be used as a poly-dT primer.

plasmid

A small DNA molecule that can replicate independently within a host cell and thus make many copies of an inserted gene is called a?

DNA

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

Cloned genes can be used in trials of gene therapy, cloned genes can be introduced into bacteria to make medicines, the expression of a closed gene can be used to discover its cellular function.

Which of the following are common uses of gene cloning?

cut the DNA backbone prior to inserting the DNA to be cloned

*restriction* *endonucleases* are used in *gene* *cloning* to?

detect and quantify the amount of a specific RNA

Reverse transcriptase PCR can be used to?

Cosmids, YACs, BACs

Select all vectors that can be used to clone large segments of DNA?

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?

recombinant

A vector that contains a piece of chromosomal DNA is referred to as a _______ vector.

genomic library

After digesting all of the chromosomal DNA of an organism with restriction enzymes and recombining the DNA into vectors, the collections of recombinant vectors resulting is called a _____ _____.

competent

Cells that can take up DNA from the medium are considered ______ cells?

to protect the cell against invasion by bacteriophages

How does a bacterial cell use restriction enzymes?

transformed cells can grow in conditions that untransformed cells cannot

How is *transformation* recognized?

lowered

In PCR, the temperature must be _______ from the denaturation temperature in order for primers to anneal.

cloning

Making many copies of a particular DNA segment using vectors or the polymerase chain reaction is called gene ____.

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 polymerases are added.

Order the steps in DNA sequencing.

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.

Order the steps in cloning a gene.

1. denaturation 2. primer annealing 3. primer extension

Order the steps in one cycle of a PCR reaction.

1. Create complementary DNA using reverse transcriptase 2. Attach linkers to cDNAs using DNA ligase. 3. Cut the cDNAs and the vectors with same restriction enzyme. 4. Ligate the cDNAs and the vectors together. 5. Transform recombinant vectors into host cells

Place in the correct order the steps in constructing a cDNA library, beginning with the first step at the top?

reverse transcriptase

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

gene cloning

The replication of recombinant DNA molecules inside a host cell is one form of?

southern blotting

The technique used to detect a particular gene sequence within a mixture of many fragments of DNA is called _____ _____.

the DNA sequence

The use of dideoxyribonucleotides with different colored fluorescent dyes allows the detection of?

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*.

Thermocycler

To perform PCR, a machine called a ________ automates the timing of each cycle.

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

What is a plasmid?

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?

western blotting

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

ligase

When cloning a gene into a vector, the sugar-phosphate backbone of each DNA molecule is covalently linked by the enzyme DNA _______.

cosmid

Which of the following vectors would you use to clone a large pieces of DNA?

taq polymerase

What is used to *amplify* *DNA* rather than RNA?

1. separate the proteins in a mixture using SDS-PAGE 2. blot the proteins onto a nylon membrane 3. incubate the nylon membrane the primary antibody. 4. incubate the nylon membrane with the secondary membrane. 5. treat the nylon mesh with a reagent that causes color changes.

Order the steps in preforming a Western blot.