Lab 7

Question 1

biotechnology

Answer 1

the use of organisms or their components to make or modify products useful to humans

Question 2

traditional biotechnology

Answer 2

• selective plant and animal breeding

- yeast in fermentation to produce cheese, bread, beer and wine

Question 3

modern biotechnology

Answer 3

-manipulation of DNA in vitro

Question 4

Each DNA strand is composed of a series of….

Answer 4

nucleotides

Question 5

Nucleotides are composed of…

Answer 5

• nitrogenous base,
• sugar (deoxyribose)
• phosphate group (negatively charged)

Question 6

Nucleotides are joined by covalent…..

Answer 6

phosphodiester bonds

Question 7

Between the two strands of DNA, ____________ are formed between ____________________.

Answer 7

• hydrogen bonds

- complementary nitrogenous bases (AT/GC)

Question 8

Biological samples collected for DNA analysis must contain ___________ cells.

Answer 8

-nucleated (bodily fluids, hair follicle w/ root attached, skin, organs, and bones)

Question 9

What are the steps in processing and analyzing DNA

Answer 9

• DNA extraction
• Polymerase Chain Reaction
• Restriction fragment analysis
• interpreting the results

Question 10

List the 3 solutions used in DNA extraction and describe them.

Answer 10

• sports drink: is isotonic to environment inside the cheek cells so cells will not burst + release DNA prior to collection
• detergent (cell lysis sol’n): lyses the plasma membrane and nuclear membrane of each cell
• ethanol: DNA is insoluble in alcohol so it precipitates out of the sol’n into the ethanol layer

Question 11

What are the 4 ingredients required for PCR

Answer 11

• DNA extract
• 4 dNTPs (dATP, dCTP, dGTP, dTTP)
• primers
• DNA polymerase (Taq polymerase)

Question 12

What are the 3 steps in each PCR cycle?

Answer 12

• DNA denaturation: heat to separate strands
• Anneal primers: cool so primers can bond to strands
• Extend primers: heat to allow DNA polymerase to add dNTPs to the end of the primers (DNA synthesis)

Question 13

What are the steps in Restriction Fragment Analysis?

Answer 13

• restriction digest

- gel electrophoresis

Question 14

Restriction Digest

Answer 14

restriction digest of the PCR product

• a restriction enzyme is added to the PCR product
• the sol’n is placed in an incubator
• enzyme cuts theDNA in the PCR product into specific fragment sizes and numbers

Question 15

The restriction enzyme EcoRI recognizes and cuts DNA within the nitrogenous base sequence….

Answer 15

GAATTC.

Question 16

restriction fragments

Answer 16

the resulting lengths of DNA from restriction digest

Question 17

gel electrophoresis

Answer 17

a process that allows us to separate the restriction fragments produced by each digested PCR product into an ordered group, based upon molecular size differences

Question 18

DNA is _______ charged

Answer 18

negatively

Question 19

The principle of gel electrophoresis is based on …

Answer 19

• the negative charge of DNA

- the differences in size of the DNA fragments

Question 20

What is the gel used in gel electrophoresis?

Answer 20

-agarose

Question 21

What stain is used in gel electrophoresis and why?

Answer 21

-Fast Blast, which can be visualized with white light

Question 22

recombinant DNA

Answer 22

when DNA from two different sources are combined into one molecule

Question 23

GMO

Answer 23

• genetically modified organism or transgenic organism

- an organism that has acquired genes through any artificial process, including recombinant DNA technology

Question 24

restriction fragment analysis

Answer 24

-process that enables an indirect comparison of nucleotide sequences in diff. amplified DNA samples

Question 25

Th gel is submerged in a chamber containing salt water because....

Answer 25

it conducts electricity

Question 26

List and describe 3 examples of modern biotechnology and how they benefit humans.

Answer 26

- insulin is manufactured by inserting the gene for insulin into a plasmid (recombinant DNA) which people take to regulate they blood sugar - cells are modified in calfs with genes for producing lysostaphin, a protein that kills staphylococcus aureus bacteria, which leads to mastitis disease in cows. - genetic modification of livestock to enable expression of selected genes in their mammary cells like important proteins for dissolving blood clots and treating hemophilia and emphysema are purified from the milk of transgenic animals. - tissue culture techniques are used to clone many different kinds of natural and transgenic plants with desirable genetic traits for use agricultural production, forestry and scientific research