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AQA A Level Biology > 8-21 DNA technology > Flashcards

8-21 DNA technology Flashcards

1
Q

What is a blunt end?

A

When DNA is cut straight through the double strand without seperating bases

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2
Q

What is the process of transformation?

A

First increase the permeability of the membranes, e.g. use calcium ions for bacteria
Keep cold to reduce metabolic activity

Once its exposed quickly raise the temperature to 42 (heat shock)
This causes the plasmid to be quickly expadited into the bacteria
This encourages the movement of the vector into the host cell

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3
Q

What are the problems that can occur during insertion?

A

The DNA fragments can join up with each other because it is looking for complimentary pairs
Sometimes plasmids close up before the gene has been inserted because the sticky ends are complementary

Sometimes two plasmids join

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4
Q

What are the advantages of recombinant DNA technology?

A

GMO plants produce plant organs which can then be harvested
Replacing defective genes can be used to cure genetic disorders

GMO crops can survive harsher conditions

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5
Q

What are the risks of recombinant DNA technology?

A

Can a company patent a gene and own DNA?
Could a new disease be created accidentally?

People could change their genetic makeup (eugenetics)

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6
Q

What are DNA probes?

A

Short single strands of DNA which contain a DNA marker

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7
Q

What are radioactively labelled probes?

A

Made up of nucleotides with the isotope 32P

Identified using an x-ray film

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8
Q

What are fluorescently labelled probes?

A

Probes which emit light

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9
Q

What is the process of DNA hybridisation?

A

Seperate the two strands of DNA

Add complimentary DNA from someone else to see if they’re complimentary

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10
Q

What is hybrid DNA?

A

A form of recombinant DNA where the complimentary parts of the strand anneal

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11
Q

What is recombinant DNA?

A

DNA from two different source

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12
Q

What does the annealing of probes do?

A

It confirms the base sequences

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13
Q

What can DNA be split up by?

A

Heat or DNA helicase

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14
Q

What is the function of PCR?

A

It amplifies your sample of DNA

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15
Q

How long is the DNA sample in PCR?

A

An entire genome or a fragment

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16
Q

What does PCR stand for?

A

Polymerase chain reaction

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17
Q

What is required for PCR in terms of DNA polymerase?

A

DNA polymerase will denature at higher temperatures

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18
Q

What is required for PCR? (without explanation)

A

DNA polymerase
pH buffer

Thermal cycle
Primers
Free nucleotides

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19
Q

What is required for PCR in terms of the thermal cycler?

A

All ingredients are put into a thermal cycler which changes the temperature

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20
Q

What are primers?

A

Short sequences of nucleotides that have a set of bases that are complementary to the DNA fragments

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21
Q

What are the three steps of PCR? (no explanation)

A

Denaturing
Annealing

Extension
This continues in a cycle

22
Q

What is the process of denaturing in PCR?

A

95 degrees celcius
The strands seperate

Hydrogen bonds break
DNA fragments, primers and taq polymerase are added

23
Q

What is the process of annealing in PCR?

A

Cooling down to 55 degrees celcius
Primers attach to each strand of DNA

Each primer covers 2+ bases
Primers prevent the strand from rejoining
Primers anneal to the DNA
This helps the taq polymerase to bind

24
Q

What is the process of extension in PCR?

A

Heated to 72 degrees celcius
Also called elongation

End up with two double strands of DNA
Free nucleotides attach
Taq polymerase joins the free nucleotides

25
Q

What antibiotics are necessary for identification? (no explanation)

A

Ampicillin, Tetracycline

26
Q

What are the two types of identification?

A

Antibiotic resistance or fluorescence

27
Q

What is replica plating?

A

Sterile velvet is placed onto the first plate to make a copy of the colonies
This is then placed onto another plate

The plates are then compared to see which colonies were killed by Ampicillin so carry the desired gene
All done in sterile conditions

28
Q

What is the stage of growth?

A

Colonies were grown in a nutrient growth

Rinse with Ampicillin at the end to kill remaining bacteria

29
Q

What role does Ampicillin play in identification?

A

The gene for ampicillin resistance is disrupted by the inserted gene fragment, any plasmid killed by ampicillin has had the DNA fragment successfully inserted

30
Q

What role does Tetracycline play in identification?

A

The gene for tetracycline is not disrupted by the DNA fragment, if the bacteria doesn’t have a plasmid it will be killed by tetracycline

31
Q

How is fluorescene used for recombinant DNA identification?

A

Add a fluorescent gene to the DNA fragment
Insert the whole thing into the vector

If the bacterium is fluorescent, the desired gene has been inserted

32
Q

What should you do when attempting to use fluorescence identification?

A

Use a large sample of DNA so that there is still a large amount of DNA left after several attempts

33
Q

What is the difference between in vivo and in vitro cloning?

A

In vivo: the bacteria do it, the process of isolation + insertion + transformation + identification + growth
In vitro: a machine does it, a polymerase chain reaction

34
Q

What are the types of isolation?

A

Conversion of mRNA to cDNA using reverse transcriptase
Using restriction endonucleases to cut fragments containing the desired gene

Creating the gene in a gene machine, based on an already known protein structure

35
Q

Bacterial artificial chromosomes

A

-

36
Q

Genetic engineering

A

-

37
Q

Phosphodiester bond

A

-

38
Q

What is amplification?

A

a number of natural and artificial processes by which the number of copies of a gene is increased “without a proportional increase in other genes

39
Q

What are bacterial artificial chromosomes?

A

an engineered DNA molecule used to clone DNA sequences in bacterial cell

40
Q

What is a clone?

A

An identical copy of a DNA sequence or entire gene; one or more cells derived from and identical to a single ancestor cell OR to isolate a gene or specific sequence of DNA.

41
Q

What is conjugation?

A

The process by which one bacterium transfers genetic material to another through direct contact. During conjugation, one bacterium serves as the donor of the genetic material, and the other serves as the recipient. The donor bacterium carries a DNA sequence called the fertility factor, or F-factor.

42
Q

What is DNA Ligase?

A

DNA ligase is a specific type of enzyme, a ligase, that facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond

43
Q

What is DNA Primer?

A

a short, single-stranded DNA sequence used in the polymerase chain reaction (PCR) technique. In the PCR method, a pair of primers is used to hybridize with the sample DNA and define the region of the DNA that will be amplified.

44
Q

What is electrophoresis?

A

A laboratory technique used to separate DNA, RNA, or protein molecules based on their size and electrical charge. An electric current is used to move molecules to be separated through a gel. Pores in the gel work like a sieve, allowing smaller molecules to move faster than larger molecules.

45
Q

What is genetic screening?

A

Genetic screening is the process of testing a population for a genetic disease in order to identify a subgroup of people that either have the disease or the potential to pass it on to their offspring.

46
Q

What is gene therapy?

A

Gene therapy replaces a faulty gene or adds a new gene in an attempt to cure disease or improve your body’s ability to fight disease.

47
Q

What are genetic markers?

A

A genetic marker is a DNA sequence with a known physical location on a chromosome. Genetic markers can help link an inherited disease with the responsible gene.

48
Q

What is genomics?

A

Genomics describes the study of all of a person’s genes (the genome).

49
Q

What are germline cells?

A

A germ line is the sex cells (eggs and sperm) that are used by sexually reproducing organisms to pass on genes from generation to generation.

50
Q

What is golden rice?

A

Golden Rice was produced by adding two genes to the genome of rice one from Erwinia uredovora which is a soil bacterium which codes for crtI (carotene desaturase) gene and another from daffodil (Narcissus pseudonarcissus) which codes for gene psy (phytoene synthase).

AQA A Level Biology (19 decks)

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