question

Autoradiograph

answer

is an image on x ray film or nuclear emulsion produced by the pattern of decay emissions from a distribution of a radioactive substance.

question

Blunt end

answer

The simplest DNA end of a double stranded molecule is called this. IN a blunt-ended molecule both strands terminate in a base pair.

question

Nick translation

answer

DNA polymerase I is used to replace some of the nucleotides of a DNA sequence with their labeled analogues, creating a tagged DNA sequence which can used as a probe

question

Nuclease

answer

is an enzyme capable of cleaving the phosphodiester bonds between the nucleotide subunits of nucleic acids.

question

T or F- RNA is easily degraded and precautions must be take during its isolation.

answer

True

question

Sedimentation by centrifugation does what?

answer

Separates macromolecules by size and shape

question

Reverse Transcriptase does what

answer

allows for the synthesis of a complementary DNA from mRNA

question

Electron Microscopy can be used how?

answer

To study macromolecules including DNA and large protein complexes

question

The process that separates particles by density is called what (centrifugation)?

answer

Equilibrium density gradient centrifugation

question

Gel Electrophoresis does what?

answer

DNAs of different size can be separated by this proceedure

question

What is used to separate small DNA molecules

answer

Polyacrylamide gels

question

What is used to separate large DNA molecules

answer

Agarose gels

question

What is used to separate very large DNA molecules

answer

Pulsed field gel electrophoresis

question

Restriction endonucleases do what?

answer

They cleave DNA at specific recognition sequences and leave blunt or sticky ends.

question

Restriction maps do what

answer

detail the placement of restriction sites and the expected sizes of fragments that will be produced upon digestion

question

What is a plasmid?

answer

an autonomously replicating small circular DNA molecule

question

What type of test is used to detect specific DNA fragments?

answer

Southern Blot

question

How can DNA fragments be cloned?

answer

In plasmid vectors that have been propagated in bacteria

question

PCR is used to

answer

exponentially amplify a DNA segment and can also be used to introduce specific site directed mutation into DNA

question

Fredrick Sanger did what

answer

devised the dideoxynucleotide method for sequencing DNA

question

The Sanger DNA sequencing method employs dideoxynucleotides for what

answer

chain termination

question

Reverse transcriptase can generate a ______ from an _____

answer

complementary DNA (cDNA) mRNA

question

cloning

answer

refers to the process of making multiple molecules. It is commonly used to amplify DNA fragments containing whole genes, but it can be used to amplify any DNA sequence such as promoters, noncoding sequences and randomly fragmented DNA.

question

palindrome

answer

example...5' TCGACT 3'
3' TCAGCT 5'

question

cohesive end

answer

refers to the properties of the end of a molecule of DNA or a recombinant DNA molecule. See sticky end

question

polymerase chain reaction (PCR)

answer

is used to amplify a single or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence

question

complementary DNA (cDNA)

answer

is DNA synthesized from a mature mRNA template in a reaction catalyzed by the enzyme reverse transcriptase and the enzyme DNA polymerase.- IT is often used to clone eukaryotic genes in prokaryotes.

question

electron microscopy

answer

process that uses a particle beam of electrons to illuminate a specimen and produce a magnified image.

question

pulsed field gel electrophoresis

answer

The procedure is similar to standard gel electrophoresis, except that the charge is periodically switched among three directions, on central and two to each side. The smaller fragments will eventually make their way towards the bottom- useful in very large fragments

question

dideoxy-terminator cycle sequencing

answer

This uses ddNTPs as DNA chain terminators. It requires a single stranded DNA template, a DNA primer, a DNA polymerase, normal dNTPs, and ddNTPS.

question

recombinant plasmid

answer

contains DNA fragment inserted within the plasmid vector and then reintroduced into living bacterial cells.

question

dideoxynucleotide triphosphate (ddNTP)

answer

are nucleotides lacking a 3'hydroxyl group on their deoxyribose sugar. The affect of this is after being added to a growing nucleotide chain, no phosphodiester bond can be created.

question

DNA chip

answer

consists of a solid surface to which DNA fragments are attached. When copies hybridize the chip shows this.

question

restriction endonuclease

answer

is an enzyme that cuts double stranded or single stranded DNA at specific recognitions nucleotide sequences known as restriction sites.

question

restriction map

answer

is a map of known restriction sites within a sequence of DNA- this can be used to engineer plasmids or other relatively short pieces of DNA and sometimes longer pieces of DNA

question

DNA polymerase I

answer

is an enzyme that participates in the process of DNA replication in prokaryotes. It removes the RNA primer, and fills in the necessary nucleotides of the okazaki fragments.

question

reverse transcriptase

answer

is a DNA polymerase enzyme that transcribes single-stranded RNA into double-stranded DNA.

question

SDS-PAGE

answer

(Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) destroys the shape of proteins so that we can measure it by size

question

genomic library

answer

is a population of host bacteria, each of which carries a DNA molecule that was inserted into a cloning vector, such that the collection of cloned DNA molecules represents the entire genome of the source organism.-Also represents the collection of all of the vector molecules, each carrying a piece of the chromosomal DNA of the organism, prior to the insertion of these molecules into the host cells.

question

bacterial artificial chromosome (BAC)

answer

is a DNA construct based on a functional fertility plasmid, used for transforming and cloning in bacteria, usually E. coli. These are often used to sequence the genome of organisms in genome projects.

question

probe

answer

is a fragment of DNA or RNA of variable length which is used in DNA or RNA samples to detect the presence of nucleotide sequences. The probe thereby hybridizes to single stranded nucleic acid whose base sequence allows base pairing.

question

expression vector

answer

is generally a plasmid that is used to introduce a specific gene into a target cell. Once the expression vector is inside the cell, the protein that is encoded by the gene is produced by the cellular transcription and translation machinery ribosomal complexes.

question

endonuclease

answer

Nucleases that act within a strand

question

sedimentation coefficient (s)

answer

It is defined as the ratio of a particle's sedimentation velocity to the acceleration that is applied to it causing sedimentation. s=Vt/a

question

equilibrium gradient centrifugation

answer

is a type of centrifugation procedure widely used in biochemistry to separate molecules based on their buoyant density. It is achieved by pinning biological preparations at high g force over long periods of time, in buffers or solutions containing a varying amount of viscous molecule.

question

site directed mutagenesis

answer

is a technique in which mutation is created at a defined site in a DNA molecule. In general it requires that the wild type gene sequence is known

question

ethidium bromide

answer

is an intercalating agent commonly used as a fluorescent tag

question

Southern Blot

answer

used to detect a specific DNA sequence in DNA samples. Combines transfer of electrophoresis separated DNA fragments to a filter membrane and subsequent fragment detection by probe hybridizaiton

question

exonuclease

answer

nucleases that act only at the end of a nucleic acid, removing a single nucleotide at a time

question

sticky end

answer

refers to the properties of the end of a molecule of DNA or a recombinant DNA molecule. When the end of a DNA has a long overhang (non base paired end) it is called this

question

gel electrophoresis

answer

The migration of charged molecules (such as nucleic acids or proteins) through an agarose or polyacrylamide gel under the influence of an electric field

question

sucrose gradient centrifugation

answer

A density gradient is created by gently overlaying lower concentrations of sucrose on high concentration. Then the protein is added and centrifuged, the proteins separate into bands according to their density and the density of succrose surround them.

question

hybridization

answer

is the process of establishing a non-covalent sequence specific interaction between two or more complementary strands of nucleic acids into a single hybrid, which in the case of two strands is referred to as a duplex.

question

svedberg (s)

answer

10^-13 seconds- s=velocity/centrifugal force

question

intercalation

answer

insertion of the planar phenanthridinium ring between adjacent base pairs cause the DNA molecule to unwind. No covalent bonds are altered and hydrogen bonds between base pairs remain intact

question

template strand

answer

or non coding strand, is used as a template for RNA synthesis. As transcription proceeds, RNA polymerase traverses the template strand and uses base pairing complementarity with the DNA template to create an RNA copy.

question

isoschizomer

answer

are pairs of restriction enzymes specific to the same recognition sequence. For example (CGTAC/G) and (CGTAC/G)

question

vector

answer

is a DNA molecule used as a vehicle to transfer foreign genetic material into another cell. The four major types of vectors are plasmids, bacteriophages and other viruses, cosmids and artificial chromosomes.

question

Klenow fragment

answer

is a large protein fragment produced when DNA polymerase I from E. coli is enzymatically cleaved by the protease subtilisin- it is useful in synthesis of double stranded DNA from Single stranded templates
-helping to create blunt ends
-digesting away protruding overhangs
-preparation of radioactive DNA probes

question

Western Blot

answer

Uses gel electrophoresis to separate, then the proteins are transfered to a membrane where they are probed using antibodies specific to the target protein.

question

Microarray

answer

is a multiplex lab on a chip. it is a 2D array on a solid substrate, that assays large amounts of biological material using high-throughput screening methods

question

two dimensional gel electrophoresis

answer

gel electrophoresis that is then separated out again except in a 90 degree angle

question

fusion protein

answer

(or chimeric proteins) are proteins created through the joining of two or more genes which originally coded for separate proteins.

question

cDNA clone

answer

a clone of the complementary DNA strand formed

question

cDNA library

answer

is a combination of cloned cDNA fragments inserted into a collection of host cells, which together constitute some portion of the transcriptome of the organism.

question

Northern Blot

answer

uses electrophoresis to separate RNA samples by size, and detection with a hybridization probe complementary to part of or the entire target sequence

question

codon

answer

tri nucleotide sequences

question

primer

answer

is a strand of nucleic acid that serves as a starting point for DNA synthesis.

question

affinity chromatography is

answer

purification technique in which a protein mixture is passed over a matrix to which ligands specific for the desired protein have been attached, so that the protein is retained on the matrix

question

Column chromatography

answer

the general term for purification technique in which a mixture of proteins is passed through a column containing a porous solid matrix

question

High performance liquid chromatography is

answer

a type of chromatography that uses columns packed with special chromatography resins composed of tiny spheres that attain a high degree of resolution, even at very fast flow rates

question

DNA synthesis always occurs from ...

answer

the 5' to 3'

question

two dimensional gel electrophoresis is

answer

technique for protein separation in which the protein mixture is run first in one direction and then in a direction at right angles to the first.

question

SDS polyacrylamide gel electrophoresis

answer

technique in which a protein mixture is separated by running it though a gel containing a detergent that binds to and unfolds the proteins

question

nuclear magnetic resonance spectroscopy

answer

analysis of the release of the electromagnetic radiation by atomic nuclei in a magnetic field due to flipping of the orientation of their magnetic dipole moments

question

(SDS-PAGE) Western Blotting, (Immunoblotting)

answer

Technique by which proteins are separated by electrophoresis, immobilized on a paper sheet, and then analyzed, usually by means of a labeled antibody

question

A virus or plasmid that carries a DNA sequence into a suitable host cell and there directs the abundant synthesis of the protein encoded by the sequence

answer

expression vector

question

Any DNA molecule formed by joining DNA segments from different sources is a

answer

recombinant DNA

question

Small circular DNA molecules that replicates independently of the genome and can be used for DNA cloning

answer

plasmid vector

question

A collection of clones that contain a variety of DNA segments from the genome of an organism

answer

genomic DNA library

question

One of a large number of enzymes that can cleave a DNA molecule at any site where a specific short sequence of nucleotides occur

answer

restriction nuclease

question

Manipulation of DNA with precision in vitro or in vivo

answer

genetic engineering

question

A DNA clone of a DNA copy of an mRNA molecule

answer

cDNA clone

question

Technique in which RNA fragments, separated by electrophoresis are immobilized on a paper sheet and then detected by hybridization with a labeled nucleic acid probe

answer

Northern Blotting

question

Technique for generating multiple copies of specific regions of DNA by the use of sequence specific primers, and multiple cycles of DNA synthesis

answer

PCR

question

Prokaryotic cloning vector that can accommodate large pieces of DNA up to 1 million base pairs.

answer

Bacterial artificial chromosome

question

The process whereby two complementary nucleic acid strands form a double helix

answer

hybridization

MMBIO 240 CH 5 EVANS – Flashcards

Unlock all answers in this set

Haven't found what you were looking for?

Search for samples, answers to your questions and flashcards