Module 3 Biotech & Bioinformatics

You determine that you have only three copies left of an important DNA fragment, so you decide to amplify it. Using flanking primers, how many PCR cycles would you have to run to generate over one billion (10⁹) copies of the fragment?

28 (3 × 2^n = 10^9)

What is the specific application of reverse transcriptase in the preparation of cDNA?What is the specific application of reverse transcriptase in the preparation of cDNA?

amplification of RNA to DNA

What is a cDNA molecule?

complementary or clone DNA

ß-galactosidase

i. lacZ

One major difference between prokaryotic and eukaryotic genes is that eukaryotic genes can contain internal sequences, called ________, that get removed in the mature message.

introns

When propagating a clone in the lambda phage, would you have more immediate success if the phage entered the lysogenic or the lytic cycle?

lytic cycle

A map of the order, overlap, and orientation of physically isolated pieces of the genome.

restriction mapping

In what way are specific DNA sequences of the template amplified in the polymerase chain reaction? In other words, how does one target the target?

flanking primers

Assume that a plasmid (circular) is 3200 base pairs in length and has restriction sites at the following locations: 400, 700, 1400, 2600. Give the expected sizes of the restriction fragments following complete digestion.

300, 700, 1000, 1200

What is the function of dideoxynucleotides in Sanger DNA sequencing?

They stop synthesis at a specific site, so the base at that site can be determined.

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

hybridizes filter-bound DNA with a DNA probe.

What is bioinformatics?

a method that uses very large national and international databases to access and work with sequence information

Clones can be of a cell, an organism, or a molecule.  What characteristic do they all have in common?

All are derived from a single ancestor

A bacterial polycistronic transcription unit is one that

contains information for more than one protein product.

A BLAST search is done to

find similar gene or protein sequences.

Which of the following enzymes is used to make complementary DNA (cDNA) from RNA?

reverse transcriptase

What do PCR, reverse transcription, and dideoxy DNA sequencing all have in common?

All produce DNA chains as a product.

What is the purpose of a cDNA library?

To produce a library of expressed genes.

Compared with eukaryotic chromosomes, bacterial chromosomes are

small, with high gene density.

Which of the below are not steps in the production of genome sequence maps:

Isolate whole chromosomes.

A section of a genome is cut with three enzymes: A, B, and C. Cutting with A and B yields a 10-kb fragment. Cutting with B and C yields a 2-kb fragment. What is the expected result from a digest with A and C, if the C site lies in between the A and B sites?

An 8-kb fragment

Some vectors such as pUC18 and others of the pUC series contain a large number of restriction enzyme sites clustered in one region. What term is given to this advantageous arrangement of restriction sites?

multiple cloning site

If a restriction enzyme cuts a circular DNA into three fragments, how many restriction sites are there in the DNA?

3

Compared with prokaryotic chromosomes, eukaryotic chromosomes are

large, linear, less densely packed with protein-coding genes, mainly organized in monocistronic units with introns.

The set of all proteins encoded by the genome is called the

proteome

It is common to use ddNTPs (dideoxyribonucleoside triphosphates) in which of the following biochemical reactions?

DNA sequencing

A principal problem with inserting an unmodified mammalian gene into a bacterial plasmid, and then getting that gene expressed in bacteria, is that

bacteria cannot remove eukaryotic introns.

What is the name of the process by which bacterial colonies (cells) are transferred from one agar plate to another, maintaining the same spatial pattern?

replica plating

This term refers to the work undertaken by large teams of researchers who, through a concerted effort, clone and sequence the DNA of a particular organism.

genome project

The first commercial production of what human enzyme led to the explosion of the biotechnology industry?

insulin

There are different challenges that exist for sequencing prokaryotic and eukaryotic genomes.  Which challenge is correctly paired with the type of genome to which it relates?

Eukaryotic: repetitive DNA

All of the following are characteristics of the genomics revolution EXCEPT_____________

Inability to understand single genes

A human gene with a disease phenotype is going to be mapped by positional cloning. Which would be the most useful for this task?

Data about the inheritance of SNP markers in families with the disease

In general, the organization of genes in bacteria is different from that in eukaryotes. In E. coli, approximately 27 percent of all genes are organized into contiguous, functionally related units containing multiple genes under coordinate control that are transcribed as a single unit. Such contiguous gene families are called

operons

The difference between a genetic screening experiment and a selection experiment is that a screening experiment involves ________, whereas a selection experiment creates conditions that ________ irrelevant organisms. 

visual examination, eliminate  

In the context of molecular genetics, reverse translation refers to

assembling a DNA sequence from an amino acid sequence.

in situ hybridization

a. chromosome spread

Another word for a “DNA chip” (microscopic spots of oligonucleotides bound to glass that can be fluorescently labelled to identify levels of expression).

microarray