TOPIC 1.7 BIOTEC TOOLS AND TECHNIQUES

Question 1

What are some historical examples of biotechnology?

Answer 1

Fermentation
Selective breeding
More recently Use of antibiotics

Question 2

major applications of biotechnology?

Answer 2

medical diagnosis
environmental energy management
animal and breeding agriculture
fermentation technology [biofuels]

Question 3

what is the purpose of pcr

Question 4

what is the purpose of restriction enzymes

Question 5

what is the purpose of gel elcetrophoresis/ DNA digestion using restriction enzymes?

Question 6

What is the purpose of genome probes?

Question 7

what is the purpose of DNA Profiling [Short tandem repeats]?

Question 8

what is the purpose of DNA sequencing [electropherogram]?

Question 9

what is the purpose of recombinant DNA?

Question 10

what is the purpose of recombinant DNA?

Question 11

What is the purpose of CRISPR?

Question 12

What is the purpose of viral vectors?

Question 13

What is the purpose of viral vectors?

Question 14

What is the purpose of protein design?

Question 15

What are some techniques of biotechnologies?

Answer 15

• Many biotechnologies rely on extracting DNA before manipulating or using it.
• Another key element is amplifying (increasing the copies) a specific sample of DNA

Question 16

what is the purpose of DNA extraction?

Answer 16

The isolation of DNA from the cells of an organism is called DNA Extraction.

Question 17

What does PCR stand for?

Answer 17

Polymerase Chain Reaction

Question 18

What is PCR?

Answer 18

• is an artificial method of replicating DNA under laboratory conditions

Question 19

What does PCR multiply?

Answer 19

used to amplify large quantities of a specific sequence of DNA from an initial minute sample

Question 20

Is the PCR process vitro or in-vitro?

Answer 20

in vitro {outside the body}

Question 21

What might PCR be used for?

Answer 21

creates multiple copies to amplify segments of DNA greatly increasing the amount of DNA available for scientists to carry out DNA analysis. Example: identifying infectious diseases.

Question 22

how is PCR used in forensics?

Question 23

How is PCR used in Evolution?

Answer 23

DNA present in hair follicles or blood can be amplified and this can lead to the identification or elimination of individuals

Question 24

How is PCR used in Evolution?

Answer 24

DNA amplified by PCR is used to determine taxonomic (evolutionary) relationships between species

Question 25

How is PCR used in Gene Therapy?

Answer 25

PCR is used to synthesise large quantities of specific genes for use in gene therapy

Question 26

What are the main three processes involved in PCR?

Answer 26

Denaturing Annealing Extensions

Question 27

What are the main components found in the PCR tube?

Answer 27

DNA sample Taq Polymerase Primers Mix Buffer Nucleotides

Question 28

What is the purpose of taq polymerase in the PCR tube?

Answer 28

Taq polymerase is the heat-stable (thermostable) DNA polymerase, it automates the repetitive step of amplifying specific DNA sequences

Question 29

What is the purpose of adding the DNA sample to the PCR tube?

Answer 29

Because in order to create multiple copies of this specific DNA, a DNA sample is required to be replicated

Question 30

What is the purpose of primers in the PCR tube?

Answer 30

Two primers, are used in each PCR reaction, the complementary sequences bind to opposite strands of the template DNA, just at the edges of the region to be copied.

Question 31

What is the definition of the primer in the PCR tube?

Answer 31

A primer, a short sequence of nucleotides ~ 20 nucleotides that provides a starting point for DNA synthesis.

Question 32

What's the purpose of free nucleotides in the PCR tube?

Answer 32

Free nucleotides to make new strands of amplified DNA.

Question 33

What occurs in the first step of PCR?

Answer 33

temperature is increased to 95 degrees which causes breaking of the hydrogen bonds between the nucleotides strands of the target DNA [DENATURING]

Question 34

What occurs in the second step of PCR?

Answer 34

the temperature is decreased to around 55 degrees which allows primers to anneal to the separated strands of target DNA through the formation of hydrogen bonds between complementary bases. [ANNEALING]

Question 35

What occurs in the third step of PCR?

Answer 35

The temperature is increased to 72 degrees which is the optimum for the DNA polymerase enzyme. DNA polymerase extends the primers and creates new nucleotide strands using free DNA nucleotides. [EXTENSIONS]

Question 36

How many steps is the PCR process and how long does it last?

Answer 36

This 3 stage PCR cycle lasts 5 minutes, and is repeated 30 to 40 times to create many copies of DNA.

Question 37

What is a quick summary of PCR step 1?

Answer 37

Step 1: the DNA is denatured through heating. This breaks the hydrogen bonds between the chains. Separating the DNA into two strands.

Question 38

What is a quick summary of PCR step 2?

Answer 38

Step 2: the mixture is cooled [annealed]. This allows for the primers to anneal/attach to each 3’ end of each strand.

Question 39

What is a quick summary of PCR step 3?

Answer 39

Step 3: The mixture is then reheated, for DNA polymerase to attach nucleotides. - Heat-tolerant DNA polymerase then replicates the region of DNA. - Takes longer for polymerisation of nucleotides

Question 40

Why is the cycles of PCR repeated?

Answer 40

Repeated cycles of heating and cooling amplify this region of DNA by thermal cycler (~30 times), to create multiple copies of the segments.