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Plant Bio > Final Lab Sections on Biotechnology > Flashcards

Final Lab Sections on Biotechnology Flashcards

1
Q

What food called when they are produced from crop plants that have had their genetic makeup altered to produce valuable or desirable traits that would not naturally occur?

A 
Genetically Modified (GM) 
or
Genetically Engineered (GE)
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2
Q

What is the rearrangement of DNA sequences within an organism and/or the methods employed to combine genes from different organisms?

A

recombinant DNA technology

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3
Q

What are plants/organisms in which DNA from one organism is introduced into and expressed in another organism?

A

Transgenic organisms

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4
Q

What are some examples of GM plant products?

A 
Roundup Ready soybeans
virus resistant papaya
corn
wheat
rice
animal feed
medicines
vaccines
fibers
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5
Q

Name 4 potential benefits of genetic modification.

A
  1. increased disease resistance
  2. productivity (biomass)
  3. hardiness
  4. feed nutritional value
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6
Q

Why are some opposed to genetically modified crops?

A

Unknown, unclear and potential risks that are associated with relatively new technologies.

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7
Q

What are we using in lab to test for the presence of transgenes?

A

Bio-Rad GMO Investigator Kit

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8
Q

What is being used to determine if the isolated DNA from the samples contains specific transgenes?

A

Polymerase Chain Reaction (PCR)

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9
Q

What are the two sequences we are testing for (that are found in about 85% of all transgenic plants)?

A
  1. A promotor from the Cauliflower Mosaic Virus (CaMV 35s)

2. A terminator gene from Agrobacterium tumefaciens called nopaline synthase (NOS)

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10
Q

What is a promoter sequence?

A

A sequence that is required to initiate or activate transcription of a transgene at a high level in plant cells.

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11
Q

Why are viral promoters desirable to use?

A

Because they are particularly strong.

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12
Q

What is a terminator sequence?

A

a sequence that is required to stop, or inactivate, transcription for a specific transgene.

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13
Q

Who invented the Polymerase Chain Reaction (PCR)?

A

Kary Mullis in 1983

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14
Q

What is the Polymerase Chain Reaction (PCR)?

A

A process in which a specific DNA region can be copied continuously, or amplified in number of copies, under a particular set of conditions.

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15
Q

How many copies can PCR produce?

A

After 25 cycles, over 33 million.
After 50 cycles, over 66 million.
Billions of copies of a single DNA sequence within a few hours.

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16
Q

What 5 components are required to make PCR happen?

A
  1. Template DNA
  2. Taq Polymerase
  3. PCR Buffer
  4. Nucleotides
  5. Primers
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17
Q

What is template DNA?

A

The DNA that you are testing to determine whether or not it contains the sequences in which you are interested.

18
Q

What is Taq Polymerase?

A

An enzyme capable of synthesizing new DNA strands, isolated from the heat-loving bacterium Thermus aquaticus.

19
Q

What is a PCR buffer?

A

Taq polymerase requires a specific buffer in which it can perform its activity.

20
Q

What are nucleotides and which ones are we using?

A

The building blocks of DNA

Adenine, Thymine, Cytosine, Guanine

21
Q

What are primers?

A

Single-stranded DNA molecules that are usually 16-30 nucleotides long and are mechanically synthesized by different companies.

22
Q

How do primers work?

A

When attempting to amplify a particular DNA sequence by PCR, individual primers are generated that precisely match with the ends of that sequence.

23
Q

What is a primer’s function?

A

Their function in the reaction is to anneal (bind) to the DNA sequences to which they are complimentary and serve as a starting point for the synthesis of new DNA strands by Taq Polymerase.

24
Q

How many primer sequences are we using?

A

4: 2 for the promoter and 2 for the terminator.

25
Q

What is the machine that we will use to run our reactions once everything is put together?

A

Thermocycler

26
Q

What DNA is extracted from the plant and food mater?

A

combined nuclear, chloroplast, and mitochondrial

27
Q

What are the 3 steps for PCR reactions?

A
  1. Denaturation
  2. Primer Annealing
  3. Extension
28
Q

What is the Denaturation stage?

A

At 94 degrees, the double stranded template DNA molecule separates (or melts, denatures) into two single-stranded molecules.

29
Q

What is the Primer Annealing stage?

A

The reaction is cooled down to about 60 degrees, where bonds are continuously reformed between single-stranded primers and the single-stranded template DNA sequence to which they are complimentary. Taq polymerase attaches at the 3’ ends of the bound primer sequence.

30
Q

What is the Extension stage?

A

At 72 degrees, the polymerase can work efficiently to add nucleotides that are complementary to the DNA template at the 3’ ends of the primer to produce a complementary strand that was elongated in a 5’ to 3’ direction.

31
Q

How many times are the PCR steps repeated?

A

30-40 times, in cycles, with a final long extension step at the end to ensure the DNA strands have been completely extended.

32
Q

How was it determined if the sequences of interest have been amplified?

A

The PCR products were resolved by agarose gel electrophoresis and visualized.

33
Q

What is Electrophoresis?

A

A technique used to separate charged molecules, like DNA, by size.

34
Q

What does DNA do in Electrophoresis?

A

DNA is a negatively charged molecule that will move from the negative cathode (-) through the gel toward the positive (+) anode.

35
Q

Which molecules will migrate further in Electrophoresis?

A

Smaller molecules will migrate further than larger molecules. Any differences in sizes of the DNA will be apparent as distinct bands.

36
Q

What is Agarose?

A

Extracted from seaweed, it is a chain of sugar molecules that forms a gel-like matrix. Purfified agarose is in a powdered form and will dissolve only in a boiling liquid.

37
Q

What buffer is used with the agarose?

A

A salt buffer that helps maintain the integrity of the DNA as it travels through the gel.

38
Q

What is used to poor the agarose solution so it will assume the shape we want as it solidifies?

A

Casting tray with a comb inserted to create small wells into which the samples are loaded.

39
Q

What is the special stain we are adding to the agar mixture?

A

GelRed from Biotium

40
Q

What does the GelRed do?

A

Adheres to the DNA fragments which will fluoresce under UV light.

41
Q

How will the size of the bands be determined?

A

By comparing them to the sizes of the bands in the molecular weight ruler (MWR).

42
Q

What is the Molecular Weight Ruler also called?

A

DNA Ladder

Plant Bio (5 decks)

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