Molecular Genetics Chapter 17

Which of the following is not key to performing the Polymerase Chain Reaction® for the amplification of a piece of DNA. a. A single stranded denatured DNA "template" b. 2 specific DNA primers c. A mixture of four deoxynucleotides and one dideoxynucleotide d. A DNA polymerase e. Repeating the temperature cycles many times

A mixture of four deoxynucleotides and one dideoxynucleotide

Southern blots probe

DNA

________ seals a nick in the DNA by creating a sugar phosphate bond between the adjacent disjointed nucleotides.

DNA ligase or topoisomerase

In each cycle of PCR what is the order of the steps (i.e. first, second and third)

Denaturation, primer annealing, extension

The methodology to clone animals such as sheep outlined in this article is essentially the same as the technology to clone DNA described earlier in the chapter.

False

Which of the following is not an essential feature or function of a plasmid cloning vector?

Origin of transfer (oriT)

cDNA can be synthesized from mRNA by using the following enzyme

Reverse transriptase

Which of the following is not an essential feature or function of a bacterial plasmid cloning vector?

Screenable marker (e.g. lacZ)

Restriction enzymes that cut between different nucleotide pairs within a palindrome on complementary DNA stands produce _______ ended DNA fragments

Sticky

Automated sequencing differs from manual sequencing in that automated sequencing:

Uses dideoxynucleotides labeled with fluorescent dyes

The technique that uses antibodies to detect proteins on a blotted gel is called a....

Western Blot

A "probe" in molecular biology is __________.

a DNA or an RNA molecule used in hybridization reactions

An E. coli plasmid used as a cloning vector must have

a bacterial origin of replication, unique restriction sites, and a selectable marker

Yeast Artificial chromosomes must have ____ to permit them to replicate in an eukaryotic host cell:

a centromere

Which is the most specific recombinant DNA library?

cDNA

In nature, the function of restriction enzymes is to

destroy phage DNA

At the end of each cycle the temperature of the PCR reaction is raised to 72°C in order to _____

elongate the primer

In the manual Sanger sequencing (chain termination), _______ sequencing reaction(s) is(are) performed.

four

For PCR, DNA can be isolated from the following source: ___.

frozen tissue, spermatic cells, fossil specimens, and preserved tissue

Recombinant DNA research is dependent on

hosts, cloning, restriction endonucleases, and vectors

Nucleic acid blotting is widely used in recombinant DNA technology. In a Southern blot one generally

hybridizes filter-bound DNA with a DNA probe.

You want to clone a fragment of DNA into a plasmid vector such as pUC18. The plasmid is 4.36 kb long and has a single restriction site for the restriction enzyme BamHI. If you digest pUC18 with BamHI, how many linear fragments will remain?

one

Western blots detect specific ____________ in a sample

proteins

Restriction enzymes cut DNA at specific palindromic segments of DNA referred to as

recognition sites

The polymerase chain reaction is a technique that __________.

selectively replicates DNA

Restriction endonucleases are especially useful if they generate "sticky" ends. What makes an end "sticky"?

single-stranded complementary tails

Which of the following DNA restriction fragments would travel the farthest distance through an agarose gel ? (if run on the same gel at the same time)

smallest: 0.5 kb

You want to close a fragment of DNA into a plasmid vector such as pUC18. The plasmid is 5 kb long and has two restriction sits for the restriction enzyme BamHI. If you digest pUC18 with BamHI, how many linear fragments will result?

two

A "YAC" is a useful

vector