DNA Manipulation

Question 1

DNA manipulation

Answer 1

using DNA in a lab for our benefit

Question 2

Gene editing

Answer 2

Process of making targeted changes to a specific section of an organism’s genome, including adding, deleting or altering specific DNA sequence

Question 3

Restriction Enzymes (endonucleases)

Answer 3

Enzymes that cut DNA at a specific location (restriction site)

Question 4

Palindrom

Answer 4

a sequence of DNA, that you can read from 5’ to 3’ and vice versa

Question 5

Polymerase

Answer 5

joins base pairs together

Question 6

Helicase

Answer 6

Unzips DNA

Question 7

Gyrase

Answer 7

Unwinds DNA

Question 8

RNA polymerase

Answer 8

makes a polymer of RNA

Question 9

DNA polymerase

Answer 9

makes a polymer of DNA

Question 10

Reverse transcriptase

Answer 10

makes cDNA from mRNA

Question 11

endonuclease

Answer 11

cuts DNA at a specific recognition sequence

Question 12

Ligase

Answer 12

Joins DNA segments with “sticky ends”

Question 13

Gene probes

Answer 13

Single-stranded DNA or RNA complementary base sequences to target DNA. Attached with fluorescent markers. DNA must denature and then anneal

Question 14

CRISPR

Answer 14

cas9 are enzymes in the cell that can cut up DNA that matches a little piece of DNA that they hold inside of themselves.

Cas9 will take the RNA transcribed from the DNA and hold the RNA in the enzyme to travel around the cell to find DNA that is complementary to the RNA.

This can be used to cut DNA at any place we want which lets us alter the genome

Question 15

Ligase

Answer 15

Opposite of endonucleases. Enzyme that can stick two single strands of DNA together to form one longer strand

Question 16

DNA amplification

Answer 16

Using the technique called polymerase chain reaction (PCR), researchers can create vast quantities of DNA identical to trace samples

Question 17

Polymerase Chain Reaction

Answer 17

Technique by which one can copy segments of a DNA sample repeatedly to form multiple copies of the same DNA for further analysis

Question 18

PCR process

Answer 18

1. mixture of DNA fragments heated to seperate strands (92 degrees)
2. temperature cool to allow primer to bind to DNA
3. temperature increased again to allow Taq DNA polymerase to start building complementary strand
4. process is repeated over and over

Question 19

Gel Electrophoresis

Answer 19

method of separating fragments of DNA, usually according to size

Question 20

Gel Electrophoresis process

Answer 20

• sheet of gel used to seperate fragments
• DNA fragments added to one end of gel
• electric current run through gel
• electric current run through gel
• DNA is negatively charged

Question 21

Plasmid

Answer 21

double-stranded circular DNA molecules separate from the bacterial chromosome. Replicate independently. Usually has its own marker gene

Question 22

Recombinant plasmid

Answer 22

plasmids that we put foreign genes into. Some plasmids may not incorporate the gene of interest because they may reseal before the gene of interest can bind. This is because the complementary sticky ends are attracted to themselves

Question 23

Transformed bacteria

Answer 23

bacteria that have incorporated the recombinant plasmid

Question 24

Electroporation

Answer 24

using electrical currents to open pores and make the membrane permeable

Question 25

heat shock

Answer 25

freezing for 1 min, heating for 1 min to open pores, more permeable to accept plasmid

Question 26

Vectors

Answer 26

a self-replicating DNA molecule used to transmit a gene from one organism to another.

Must have the following properties:

• Be able to replicate inside their host organism
• Have one or more sites at which a restriction enzyme can cut
• Have genetic marker that allows them to be easily identified

Question 27

DNA Sequencing

Answer 27

To identify the sequence of DNA

Question 28

DNA Sequencing process

Answer 28

• DNA strand is heated, hydrogen bonds between bases break
• DNA is allowed to cool allowing primer to bind
• DNA polymerase builds complementary strands
• repeat for all four nucleotides

Question 29

DNA profiling

Answer 29

To identify DNA from different individuals

Question 30

DNA profiling process

Answer 30

small tandem repeats: Sites in the DNA where many copies of a short DNA sequence are joined end to end

• when cut with the same endonuclease, DNA fragments will be different lengths
• gel electrophoresis shows different lengths of DNA fragments due to STR

Question 31

Gene cloning

Answer 31

Making multiple copies of a gene using prokaryotes.

Question 32

Gene cloning process

Answer 32

• endonuclease that leave sticky are used to cut before and after a gene
• gene fragment is inserted into another species for cloning

Question 33

Genetic Screening

Answer 33

Refers to an organised program of testing of groups or a population to achieve early detection or exclusion of inherited disorders. Testing someone for the presence of an allele known to cause disease.

Question 34

Positive results of genetic screening

Answer 34

• Can adjust lifestyle to prevent the condition
• Early treatment prevention
• Planning for life

Question 35

Negative results of genetic screening

Answer 35

• False positive results
• Discrimination for jobs, insurance
• Costs, resources used in testing