Chapter 5: Molecular Tools for Studying Genes

Question 1

sodium dodecyl sulfate (SDS)

Answer 1

denatures proteins and adds negative charge to proteins so they’ll migrate toward the anode

Question 1

used to determine major landmarks within a strand or segment of DNA: a physical map of locations of restriction sites within piece of DNA

Answer 1

restriction mapping

Question 2

restriction mapping

Answer 2

used to determine major landmarks within a strand or segment of DNA: a physical map of locations of restriction sites within piece of DNA

Question 2

aka size exclusion; large proteins move faster b/c smaller ones get stuck in the matrix

Answer 2

gel filtration chromatography

Question 3

a technique used to separate/purify fragments/strannds of nucleic acids

always includes known DNA ladder

Answer 3

gel electrophoresis

Question 5

gel electrophoresis

Answer 5

a technique used to separate/purify fragments/strannds of nucleic acids

always includes known DNA ladder

Question 6

hybridization

Answer 6

the ability of a strand of DNA/RNA to form a double helix with another strand of DNA/RNA

Question 7

used to separate/purify polypeptides

Answer 7

polyacrylamide gel electrophoresis (PAGE)

Question 7

invented by Edward Southern; transferred DNA from gel to thin membrane of microcellulose

gel is run and placed in “sandwich” with membrane and exposed to current to transfer DNA. Membrane/filter is then washed, blocked, and exposed to highly specific probe

Answer 7

southern blotting

Question 8

a powerful tool used for forensics, paternity testing, medical screening, or phylogeny.

A HaeIII restriction cut of a specific region of DNA followed by Southern blot

Answer 8

fingerprinting

Question 9

placing (loading) DNA/RNA into agarose gel and subjecting to current, causing DNA to migrate toward positive pole

Answer 9

DNA electrophoresis

Question 10

column chromatography

Answer 10

method for fractionating nucleic acids or proteins using stationary phage from cellular lysate (i.e. nucleic acids in solution)

Question 10

tracers, labels, probes

Answer 10

highly sensitive techniques used to detect a gene, nucleic acid, or protein of interest

Question 11

DNA electrophoresis

Answer 11

placing (loading) DNA/RNA into agarose gel and subjecting to current, causing DNA to migrate toward positive pole

Question 12

nucleic acids with more negative charge migrate more slowly through the matrix

Answer 12

Ion-exchange chromatography

Question 12

specific region of DNA

Answer 12

minisatellite

Question 14

phosphorimaging and light detect (chemiluminescence)

Answer 14

tools to evaluate strength of molecular signal (from radioactive or chemical tracers)

Question 14

denatures proteins and adds negative charge to proteins so they’ll migrate toward the anode

Answer 14

sodium dodecyl sulfate (SDS)

Question 15

polyacrylamide gel electrophoresis (PAGE)

Answer 15

used to separate/purify polypeptides

Question 16

Ion-exchange chromatography

Answer 16

nucleic acids with more negative charge migrate more slowly through the matrix

Question 17

the ability of a strand of DNA/RNA to form a double helix with another strand of DNA/RNA

Answer 17

hybridization

Question 18

highly sensitive techniques used to detect a gene, nucleic acid, or protein of interest

Answer 18

tracers, labels, probes

Question 20

southern blotting

Answer 20

invented by Edward Southern; transferred DNA from gel to thin membrane of microcellulose

gel is run and placed in “sandwich” with membrane and exposed to current to transfer DNA. Membrane/filter is then washed, blocked, and exposed to highly specific probe

Question 21

detection of RNA

blotted from gel to thin membrane of nitrocellulose, then probed with highly specific probe (usually cDNA) for specific RNA of interest

e.g. rat tissue probed for glyceraldehyde-3-phosphate dehydrogenase (G3PHD) [helps make ATP in cells and tissures]

Answer 21

Northern blotting (e.g.)

Question 22

method for detecting radioactive (P32, S35, C14) or illuminating compounds (chemiluminescence) that bind molecules of interest. Gel is exposed to X-ray film & electrons from gel/membrane pass through film to take a picture of itself

Answer 22

autoradiography

Question 24

autoradiography

Answer 24

method for detecting radioactive (P32, S35, C14) or illuminating compounds (chemiluminescence) that bind molecules of interest. Gel is exposed to X-ray film & electrons from gel/membrane pass through film to take a picture of itself

Question 25

method for fractionating nucleic acids or proteins using stationary phage from cellular lysate (i.e. nucleic acids in solution)

Answer 25

column chromatography

Question 26

column chromatography

Answer 26

method for fractioning/separating nucleic acids or proteins using a stationary phase from a cellular lysate (nucleic acids in solution)

Question 28

minisatellite

Answer 28

specific region of DNA

Question 29

method for fractioning/separating nucleic acids or proteins using a stationary phase from a cellular lysate (nucleic acids in solution)

Answer 29

column chromatography

Question 30

Northern blotting (e.g.)

Answer 30

detection of RNA

blotted from gel to thin membrane of nitrocellulose, then probed with highly specific probe (usually cDNA) for specific RNA of interest

e.g. rat tissue probed for glyceraldehyde-3-phosphate dehydrogenase (G3PHD) [helps make ATP in cells and tissues]

Question 32

fingerprinting

Answer 32

a powerful tool used for forensics, paternity testing, medical screening, or phylogeny.

A HaeIII restriction cut of a specific region of DNA followed by Southern blot

Question 33

gel filtration chromatography

Answer 33

aka size exclusion; large proteins move faster b/c smaller ones get stuck in the matrix

Question 34

tools to evaluate strength of molecular signal (from radioactive or chemical tracers)

Answer 34

phosphorimaging and light detect (chemiluminescence)