Booklet 14 Flashcards Preview

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Flashcards in Booklet 14 Deck (95):
1

DNA manipulation alters an organisms (...) either by (...) (...) DNA or (...) (...) DNA

DNA manipulation alters an organisms DNA either by adding new DNA or editing the existing DNA

2

Genetic engineering

refers to the scientific method for the artificial manipulation of genes

3

Genetic engineering builds on traditional methods of genetic manipulation that includes (...) programmes and the deliberate (...) of (...) by (...) organisms to (...) (particularly plants)

Genetic engineering builds on traditional methods of genetic manipulation that includes artificial breeding programmes and the deliberate introduction of novel traits by exposing organisms to mutagens (particularly plants)

4

genetic engineers require (...) to (...) , (...) , (...) , (...) and (...) DNA

genetic engineers require tools to cut, edit, join, copy and seperate DNA

5

restriction enzymes also known as (...) is a tool used by genetic engineers to (...) DNA

restriction enzymes also known as endonuclease is a tool used by genetic engineers to cut DNA

6

restriction enzymes have the ability to (...) DNA molecule at (...) sequences of (...) called (...). once (...) the (...) binds to the DNA and (...) the DNA in a (...) and (...) way

restriction enzymes have the ability to cut DNA molecule at very precise sequences of 4 to 8 base pairs called recognition sites. once recognised the enzyme binds to the DNA and cuts the DNA in a fixed and predictable way

7

restriction enzymes (...) in (...) , thought to have (...) as a d (...) against (...)

restriction enzymes occur naturally in bacteria, thought to have evolved as a defence mechanism against viruses

8

there are over (...) types of restriction enzymes that recognise about (...) different recognition sites

there are over 400 types of restriction enzymes that recognise about 100 different recognition sites

9

restriction enzymes are named after the (...)from which they have been (...)

restriction enzymes are named after the bacterial species from which they have been isolated

10

the position where a (...) enzyme can (...) is it (...) and is where a particular (...) of (...) occur

the position where a cutting enzyme can snip is it recognition sequences and is where a particular order of nucleotides occur

11

some restriction enzymes cut the (...) strands of a DNA molecule at points (...) to produce (...) that are (...)

some restriction enzymes cut the two strands of a DNA molecule at points directly opposite each other to produce cut ends that are blunt

12

other cutting enzymes cut (...) strand at (...) point, but cut the (...) strand at a point that is (...), the (...) cut ends made by these cutting enzymes are called (...). these sticky ends are (...)

other cutting enzymes cut one strand at one point, but cut the other strand at a point that is not directly opposite, the overhanging cut ends made by these cutting enzymes are called sticky. these sticky ends are complementary

13

EcoRI (...) the (...) at a (...) sequences, which leaves (...) ends called (...) ends

dna cut in such a way produces ends, which may only be joined to other (...) ends with a (...)

EcoRI cuts the DNA at a specific base sequences, which leaves overhanging ends called sticky ends

dna cut in such a way produces ends, which may only be joined to other sticky ends with a complementary base sequence

14

AluI cuts the (...) at a (...) sequence which leaves (...) ends called (...) ends.

dna cut in such. way is able to be joined to (...) end but tend to be (...)

AluI cuts the dna at a specific base sequence which leaves no overhanging ends called blunt ends.

dna cut in such. way is able to be joined to other blunt end fragments but tend to be non-specific

15

CRISPR-Cas9 is a (...) compromising (...)and (...)

CRISPR-Cas9 is a complex compromising Cas9 endonuclease and sgRNA

16

CRISPR complex (...) DNA at (...) sequences and can be used to (...) genes

CRISPR complex cuts DNA at very specific sequences and can be sued to edit genes

17

CRISPR-Cas9 is an endonuclease complex occurring (...), which use it to (...) of (...)

CRISPR-Cas9 is an endonuclease complex occurring naturally in bacteria, which use it to chop the DNA of invading viruses

18

two key components of CRISPR to work, an (...), (...), that (...) and (...) to the (...) of dna and the Cas9 endonuclease that (...) and (...) the dna

thus technology has the potential to (...) responsible for (...) diseases, (...) faulty genes (...), (...) (...) genes to an organism or to (...) the (...) of (...) genes. also it allows for more (...) and (...) gene editing at much lower (...) than previous tech.

two key components of CRISPR to work, an RNA guide, sgRNA, that locates and binds to the target piece of dna and the Cas9 endonuclease that unwinds and cuts the dna

thus technology has the potential to correct mutations responsible for genetic diseases, switch faulty genes off, add new genes to an organism or to study the effects of specific genes. also it allows for more precise and efficient gene editing at much lower cost than previous tech.

19

Gene knocking in or gene (...) refers to DNA sequences (...) into the DNA break. for example it allows a (...) sequence to be (...) with the (...) sequence to (...) (...) function

Gene knocking in or gene editing refers to DNA sequences inserted into the DNA break. for example it allows a faulty gene sequence to be replaced with the correct sequence to restore normal gene function

20

gene knock out or gene (...) refers to as the cells (...) mend the (...), (...) occur resulting in the (...) or (...) of (...). the resulting (...) mutation changes the way the (...) is read either disabling (...) or producing a (...). this technique can be used to silence a faulty gene

gene knock out or gene silencing refers to as the cells normal repair process mend the broken dna, errors occur resulting in the insertion or deletion of nucleotide bases. the resulting frame shift mutation changes the way the nucleotide sequence is read either disabling gene function or producing a STOP signal. this technique can be used to silence a faulty gene

21

Cas9 is (...) to the (...) by (...). (...) (...) the dna and (...) both (...) at a (...) point

Cas9 is guided to the target site by sgRNA. Cas9 unwinds the dna and cuts both strands at a specific point

22

sgRNA is a short (...) sequence designed to (...) (...) to the (...). it contains a (...) section which is (...) to the DNA of interest

sgRNA is a short synthetic sequence designed to guide Cas9 to the site of interest. it contains a nucleotide section which is complementary to the DNA of interest

23

PAM sequence lies directly (...) of the (...) of the (...) DNA strand, (...) of PAM by (...) (...) the DNA allowing the (...) to be (...). Cas9 will not (...) if PAM is (...)

PAM sequence lies directly downstream of the target sequence of the non target DNA strand, recognition of PAM by Cas9 destabilise the DNA allowing the sgRNA to be inserted. Cas9 will not function if PAM is absent

24

dna fragments can be (...) using the technique of electrophoresis as DNA is (...) charged due to their (...) in their (...)

dna fragments can be sorted using the technique of electrophoresis as DNA is negatively charged due to their phosphate groups in their sugar-phosphate backbone

25

electrophoresis sorts dna fragments according to their (...)

electrophoresis sorts dna fragments according to their size

26

electrophoresis:
fragments of DNA are placed in a jelly like (...)supporting material known as a (...). the gel is then (...) to an (...) with the positive pole at the (...) end and the negative pole at the (...) end

fragments of DNA are placed in a jelly like supporting material known as a gel. the gel is then exposed to an electric field with the positive pole at the far end and the negative pole at the starting end

27

electrophoresis:
standards consist of dna fragments of (...) and are used to compare the (...)of the sample dna

standards consist of dna fragments of known length and are used to compare the size of the sample dna

28

electrophoresis

the (...) of the unknown dna band is identified by comparing the (...) the (...) of (...) found in the standards

the size of the unknown dna band is identified by comparing the distance moved against the fragments of known size found in the standards

29

electrophoresis

the size of dna is measured in (...)

the size of dna is measured in base pairs

30

electrophoresis

dna moves from (...)terminal to (...) terminal because dna is negatively charged and is attracted to the (...) charge at the (...) terminal

dna moves from negative terminal to positive terminal because dna is negatively charged and is attracted to the positive charge at the positive terminal

31

electrophoresis

the (...) DNA fragments move most quickly and are found (...) from the starting point. the longest fragments move most (...) and are found (...) to the starting point, fragments of the same (...) move at the same (...)

the shortest DNA fragments move most quickly and are found further away from the starting point. the longest fragments move most slowly and are found closer to the starting point, fragments of the same size move at the same rate

32

electrophoresis

dna fragments are (...) so after the gel run is complete, the (...) must be (...) either through the use of a (...) or a (...) probe.

one technique makes use of the dye (...), which binds to the (...) (...) molecules.

when illuminated by (...) light the (...) bound to (...) fluoresces (...)

dna fragments are invisible so after the gel run is complete, the seperated dna bands must be made visible either through the use of a dye or a radioactive probe.

one technique makes use of the dye EtBr, which binds to the major groove of dna molecules.

when illuminated by UV light the dna bound to EtBr fluoresces pale pink

33

the end result of electrophoresis is a (...) at different (...) down the drain

the end result of electrophoresis is a series of parallel bars of dna fragments at different distances down the drain

34

a probe is a (...) strand of (...) or (...) with a base sequence that is (...) to the base sequence in one of the strands of the (...) DNA

a probe is a single strand of DNA or RNA with a base sequence that is complementary to the base sequence in one of the strands of the target DNA

35

a probe is labelled with a radioactive or fluorescent marker so that the (...) of the probe and hence the(...) can be seen

a probe is labelled with a radioactive or fluorescent marker so that the location of the probe and hence the target DNA can be seen

36

probes

fluorescent dye tag shows up as (...) when gel is (...) to (...)

fluorescent dye tag shows up as fluorescent bands when gel is exposed to UV light source

37

probes

radioactive tag shows up as a(...) band when the gel is exposed to (...)

radioactive tag shows up as a dark band when the gel is exposed to photographic film

38

probes

target dna strand has a (...) sequence that is being (...) for by the (...)

target dna strand has a complementary sequence that is being searched for by the probe

39

gene probes may be used to (...) for the (...) of a (...) on a (...), the (...) of an (...) of a (...) associated with a genetic disease or the (...) of a person to tell them apart form others ie. paternity testing

gene probes may be used to search for the position of a gene on a chromosome, the presence of an allele of a specific gene associated with a genetic disease or the genetic fingerprint of a person to tell them apart form others ie. paternity testing

40

Southern Blotting: probe use
1. dna of interest is (...) from living or dead organism (or found at crime scene)
2. (...) using (...), leaving thousands of fragments of (...)
3. dna is (...) using (...)
4. dna fragments (...) by (...) to (...) and is made (...) and (...) in place on membrane
5. when (...) finds (...) sequence of bases it will (...) to the other strand ie. (...). ((...) is the (...) between (...) stranded (...)

Southern Blotting: probe use
1. dna of interest is extracted from living or dead organism (or found at crime scene)
2. cut using restriction enzymes, leaving thousands of fragments of different sizes
3. dna is seperated using gel electrophoresis
4. dna fragments transferred by blotting to membrane and is made single stranded and fixed in place on membrane
5. when probe finds complementary sequence of bases it will join to the other strand ie. hybridisation. (hybridisation is the pairing between single stranded complementary dna segments

41

ligation

dna fragments produced using restriction enzymes may be joined by a process call ligation

42

enzyme dna ligase (...) the (...) of pieces of (...) stranded dna at their (...). the joining can produce (...) piece of dna or a (...) of dna.

two pieces of dna of (...) ie. from two different species can be (...) together using (...) and (...) to produce(...) dna

enzyme dna ligase catalyses the joining of pieces of double stranded dna at their sugar phosphate backbones. the joining can produce one longer piece of dna or a circular molecule of dna.

two pieces of dna of different origins ie. from two different species can be joined together using restriction enzymes and dna ligase to produce recombinant dna

43

ligation:

suppose that a team of genetic engineers wants to insert copies of dna fragment from a plant cell into a bacterial cell. (...) or (...) can (...) dna (...) into (...).

one examples of a vector is a (...) molecule known as a (...), which can be found in many (...) cells

suppose that a team of genetic engineers wants to insert copies of dna fragment from a plant cell into a bacterial cell. special carriers or vectors can transport dna fragments into cell.

one examples of a vector is a small circular dna molecule known as a plasmid, which can be found in many bacterial cells

44

bacterial plasmids are used in (...) dna tech

bacterial plasmids are used in recombinant dna tech

45

ligation:

plasmids are (...) pieces of dna which are able to (...) (...) of the (...) cell's (...).

plasmids are small circular pieces of dna which are able to replicate independently of the bacterial cell's chromosome.

46

ligation:

plasmids are often present in (...) copies and also often carry (...) markers.

plasmids are often present in many copies and also often carry antibiotic resistant markers.

47

Ligation process:
1. (...) pieces of dna are (...) using the (...) producing the (...)
2. (...): (...) (...) come together and join by (...). The (...) are (...) to each other by weak (...) bonds. Annealing (...) the molecules so they can be (...)
3. (...) of dna are (...) by (...) producing a (...). dna ligase (...) the (...) bond between the sugar and the phosphate group in the backbone of the dna molecule

Ligation process:
1. two pieces of dna are cut using the same restriction enzyme producing the same sticky ends
2. Annealing: two matching sticky ends come together and join by complementary base pairing. The fragments are attracted to each other by weak hydrogen bonds. Annealing stabilises the molecules so they can be permanently annealed
3. fragments of dna are permanently joined together by dna ligase producing a recombinant dna. dna ligase catalyses the phosphodiester bond between the sugar and the phosphate group in the backbone of the dna molecule

48

recombinant plasmid:

plasmid that contains dna from two or more different sources

49

methods of making specific dna fragments

1. synthesis dna from nucleotide building blocks
2. make a copy of dna using an mRNA sequence

50

method of making specific dna fragments 1
- this method makes use of an instrument called a (...)
- the base sequence of the (...) must be (...)
- dna synthesisers can (...) nucleotide sub units in a (...) to produce dna segments with (...) greater than (...) bases
- the chemical synthesis of DNA does not require a (...) nor does it require the (...)
- products can be used as (...) or (...)

method of making specific dna fragments 1
- this method makes use of an instrument called a dna syntheisiser
- the base sequence of the required dna must be known
- dna synthesisers can join nucleotide sub units in a predefined order to produce dna segments with lengths greater than 100 bases
- the chemical synthesis of DNA does not require a template strand nor does it require the enzyme DNA polymerase
- products can be used as primers or probes

51

method of making specific dna fragments 2
1. (...) of a gene from eukaryotic organism containing (...)
2. as (...), process of gene (...) creates (...) molecule
3. (...) removed by (...) forming (...) that codes for single protein
4. mRNA (...) from cell and (...)
5. (...) added which (...) (...) stranded dna molecule (...) to (...)
6. (...) dna strand made by using (...) as (...) and (...) (...)

method of making specific dna fragments 2
1. double stranded dna of a gene from eukaryotic organism containing introns
2. as normal part of cell, process of gene transcription creates primary RNA molecule
3. introns removed by restriction enzyme forming mature mRNA that codes for single protein
4. mRNA extracted from cell and purified
5. reverse transcriptase added which synthesises single stranded dna molecule complementary to mRNA
6. second dna strand made by using first as template and adding enzyme RNA polymerase

52

method of making specific dna fragments 2:
Why remove introns?
- DNA made (...), therefore (...) plasmid insertion
- (...) amount of (...) not (...) by PCR
- allows bacterial (...) to (...) (...) human gene from (...) DNA

method of making specific dna fragments 2:
Why remove introns?
- DNA made shorter, therefore easier plasmid insertion
- large amount of non coding DNA not made by PCR
- allows bacterial enzymes to properly translate human gene from reassembled DNA

53

PCR

dna amplification

54

PCR:
- using technique of PCR, researchers are ablate create larger quantities of (...) ie. (...) of DNA are (...)

PCR:
- using technique of PCR, researchers are able to create larger quantities of trace amount of DNA ie. small quantities of DNA are amplified

55

PCR:
trace DNA sources are:
1. (...) eg. single root hair, (...), drinking glass
2. (...)eg. (...)
3. (...): (...), foetal cells obtained from amniotic fluid

PCR:
trace DNA sources are:
1. crime scene eg. single root hair, dried blood stain, drinking glass
2. extinct animals eg. fragments of dna from long extinct animal
3. medical: single virus particle, foetal cells obtained from amniotic fluid

56

PCR:
process is (...), (...) and (...) and (...) using (...)

PCR:
process is cheap, quick and accurate and completed using thermal cycler

57

PCR materials

- target sequence of dna
- free dna nucleotides
- primers (short segments of single stranded dna)
- dna polymerase enzyme (TAQ polymerase)

58

PCR STEPS:
1. (...): when (...) to (...), (...) between bases break and dna (...) into sin (...)gle strands
2. (...): mixture (...) to (...). (...) added and pair with regions at (...) (...)
3. (...): mixture (...) to (...). (...) uses primers as (...) and (...) so two complete strands form. (...) must be available

PCR STEPS:
1. Denaturation: when heated to 94, weak H bonds between bases break and dna dissociates into single strands
2. annealing: mixture cooled to 55. primers added and pair with regions at either end of dna region of interest
3. exstention: mixture heated to 72. DNA polymerase enzyme uses primers as starting point and extends so two complete strands form. dna nucleotides must be available

59

PCR:
- about (...) minutes required for each PCR cycle. over time, (...) growth of dna from two original strands occur. therefore after (...) cycles, (...) copies dna exist

PCR:
- about five minutes required for each PCR cycle. over time, exponential growth of dna from two original strands occur. therefore after 20 cycles, one million copies dna exist

60

transforming bacteria:
plasmids: (...) of dna
- a plasmid is a (...) because it contains a point of (...), a base sequence where (...) begins

transforming bacteria:
plasmids: vectors of dna
- a plasmid is a self replicating dna molecule because it contains a point of origin, a base sequence where dna replication begins

61

transforming bacteria:
plasmids: vectors of dna
all vectors must have the following properties:
- be able to (...) inside the (...) organism
- have one or more (...)at which a (...) can cut
- have some kind of (...) that allow them to be easily (...)

organisms such as (...) and (...) also have dna which can be used for a vector

transforming bacteria:
plasmids: vectors of dna
all vectors must have the following properties:
- be able to replicate independently inside the host organism
- have one or more sites at which a restriction enzyme can cut
- have some kind of genetic markers that allow them to be easily identified

organisms such as viruses and yeast also have dna which can be used for a vector

62

bacterial transformation:
- in the laboratory, scientists commonly use a (...) as the carrier or vector to (...) new genes into bacterial cells
- WHEN (...) DNA IS (...) FROM AN (...) SOURCE BY (...) CELLS, THOSE (...) ARE SAID TO BE (...)

bacterial transformation:
- in the laboratory, scientists commonly use a plasmid as the carrier or vector to introduce new genes into bacterial cells
- WHEN FOREIGN DNA IS DIRECTLY TAKEN UP FROM AN EXTERNAL SORUCE BY BACTERIAL CELLS, THOSE BACTERIA ARE SAID TO BE TRANSFORMED

63

Bacterial transformation process:
1. (...) (...) from lab culture grown cell
2. (...) (...) (...) from bacterial cell
3. human dna and plasmid both treated (...) producing (...)
4. (...) cuts plasmid dna at (...) (...) tetracycline resistance gene
4. dna fragments mixed together and (...) (...) to each other by (...). (...) added to bond sticky ends
5. (...) introduced bacterial cell by adding DNA to bacterial culture. under right conditions, (...) occurs where bacteria take up plasmid from solution

Bacterial transformation process:
1. dna fragment isolated from lab culture grown cell
2. plasmid vector isolated from bacterial cell
3. human dna and plasmid both treated with same restriction enzyme producing identical sticky ends
4. restriction enzyme cuts plasmid dna at single recognition sequence disrupting tetracycline resistance gene
4. dna fragments mixed together and complementary base sticky ends attracted to each other by complementary base pairing. DNA ligase added to bond sticky ends
5. recombinant plasmid introduced bacterial cell by adding DNA to bacterial culture. under right conditions, transformation occurs where bacteria take up plasmid from solution

64

Antibiotic resistance markers
- plasmid often carries (...), ampicillin and tetracycline giving (...)
- restriction enzyme binds to tetracycline resistance gene section (...) thereby making bacteria (...) to tetracycline antibiotic
- as such, bacteria (...) in such contact

Antibiotic resistance markers
- plasmid often carries two resistant genes, ampicillin and tetracycline giving antibiotic resistance
- restriction enzyme binds to tetracycline resistance gene section disrupting function thereby making bacteria vulnerable to tetracycline antibiotic
- as such, bacteria dies in such contact

65

Getting plasmids into bacterial cells: 1.
(...): bacterial cells briefly placed in electric field that (...) them and appears to create (...) in their (...) so (...) entry (...)

Getting plasmids into bacterial cells:
electroporation: bacterial cells briefly placed in electric field that shocks them and appears to create holes in their plasma membranes so plasmid entry facilitated

66

Getting plasmids into bacterial cells: 2.
(...) bacterial cell suspending them in(...) and transferring them to (...) for (...). this increases (...)of bacterial cells thus increasing (...) of plasmid uptake

Getting plasmids into bacterial cells: 2.
heat shock bacterial cell suspending them in ice cold salt solution and transferring them to 42 deg. for less than one minute. this increases plasma membrane fluidity of bacterial cells thus increasing chance of plasmid uptake

67

isolation of transformed bacteria:
- bacterial cells that have taken up recombinant plasmid must be (...)
- not all bacteria are (...) when recombinant plasmids added to bacterial culture. some bacteria (...) plasmid, and (...) taken up are (...)

isolation of transformed bacteria:
- bacterial cells that have taken up recombinant plasmid must be isolated
- not all bacteria are transformed when recombinant plasmids added to bacterial culture. some bacteria not take up plasmid, and not all plasmids taken up are recombinant

68

transforming bacteria 3 outcomes:

transforming bacteria 3 outcomes:
1. bacteria that haven't taken up plasmid
2. bacteria that have taken up recombinant plasmid ie. transformed bacteria
3. bacteria that have taken up non-recombinant plasmid

69

transforming bacteria:
(...) of bacteria that carry recombinant plasmid can be identified by(...)- (...) to ampicillin but (...) to tetracycline

transforming bacteria:
colonies of bacteria that carry recombinant plasmid can be identified by differential antibiotic response- resistant to ampicillin but sensitive to tetracycline

70

isolation of transformed bacteria process:
- (...) pressed against(...) thereby (...) colonies of bacteria to paper
- filter paper then pressed against against (...) (or whatever the foreign gene affected)- colonies that (...) have the (...) tetracycline resistance gene

isolation of transformed bacteria process:
- filter paper pressed against agar plate thereby transferring colonies of bacteria to paper
- filter paper then pressed against against agar containing tetracycline (or whatever the foreign gene affected)- colonies that do not grow have the foreign gene disrupting tetracycline resistance gene

71

gene cloning:
- (...) occurs when the bacterium, with the recombinant plasmid, is allowed to (...) and (...) the gene
- once inside the bacteria cell, the (...) and its (...)ie. the recombinant plasmid (...) to (...) many copies
- after about (...) minutes, the bacterial cell itself divides by (...) to produce(...)
- bacteria continue to (...) along with the (...) and the (...), making millions of copies of bacterial cells and human dna

gene cloning:
- actual gene cloning process occurs when the bacterium, with the recombinant plasmid, is allowed to multiply and express the gene
- once inside the bacteria cell, the plasmid and its foreign dna ie. the recombinant plasmid multiply to produce many copies
- after about 20 minutes, the bacterial cell itself divides by binary fission to produce two daughter cells
- bacteria continue to multiply along with the plasmid and the human dna, making millions of copies of bacterial cells and human dna

72

gene cloning:
- once the bacteria, carrying the human gene has been (...) the human gene can be (...) producing its (...)
- certain proteins used in medical treatments can be produced by this (...) in (...) at (...) and (...)
- eg. (...), (...)

gene cloning:
- once the bacteria, carrying the human gene has been isolated the human gene can be switched on producing its protein product
- certain proteins used in medical treatments can be produced by this recombinant technology in large quantities at low cost and higher degree of purity
- eg. humilin, Puregon

73

gene cloning:
- once a cell line has been identified in small scale tests having giving a (...), it is moved to large scale mass culturing
- the recombinant bacteria are cultured in (...) that have a capacity of up to 50000 L

gene cloning:
- once a cell line has been identified in small scale tests having giving a high level of expression of the desired gene, it is moved to large scale mass culturing
- the recombinant bacteria are cultured in large stainless steel fermentation tanks that have a capacity of up to 50000 L

74

gene cloning:
- the culture medium kept at (...) temp. supplies all the (...) required for bacterial cell growth, wiht the bacteria (...) about every (...) minutes
- each of the million so bacterial cells in the mass culture is like a mini protein producing factory as it expresses the cloned genes carried in its recombinant plasmids

gene cloning:
- the culture medium kept at optimal temp. supplies all the requirements required for bacterial cell growth, wiht the bacteria replicating about every 20 minutes
- each of the million so bacterial cells in the mass culture is like a mini protein producing factory as it expresses the cloned genes carried in its recombinant plasmids

75

gene cloning:
- recombinant dna tech. replaced the former means of (...) that involved (...) of these (...) from (...) from(...) or (...)
- extraction from cadavers involved risk of transmission of (...) or (...)

gene cloning:
- recombinant dna tech. replaced the former means of production that involved extraction of these proteins from tissue samples from living donors or cadavers
- extraction from cadavers involved risk of transmission of viruses or prions

76

genetic screening

the testing of individuals to detect those with the alleles responsible for particular genetic disorders

77

in australia all new born babies are genetically screened for four genetic disorders for which early detection can reduce the severity of the disease:
1. (...)
2. (...)
3. (...)
4. (...)

in australia all new born babies are genetically screened for four genetic disorders for which early detection can reduce the severity of the disease:
1. phenylketonuria
2. galactosemia
3. congenital hypothyroidism
4. cystic fibrosis

78

adult screening for increased risk of disease
- genetic test can be don based on (...), to identify if an individual has a (...) known to cause a particular disease eg. (...) and (...) genes that cause (...) and (...)

some women who (...) for breast cancer genes make the decision to have a double (...) and have their (...) removed. this is quite drastic step and some people believe with the (...) that it is (...) to take such drastic precautions

adult screening for increased risk of disease
- genetic test can be don based on family medical history, to identify if an individual has a inherited gene known to cause a particular disease eg. BRCA1 and BRACA2 genes that cause breast cancer and ovarian cancers

some women who test positive for breast cancer genes make the decision to have a double mastectomy and have their ovaries removed. this is quite drastic step and some people believe with the reliability of screening that it is not necessary to take such drastic precautions

79

carrier detection

to identify people who carry a (...) that can be passed into offspring. they themselves (...) but there is (...) of the disease eg. haemophilia

allows people to make (...) regarding offspring. it gives information on the (...) of having (...) with a (...)

carrier detection

to identify people who carry a mutant allele that can be passed into offspring. they themselves show no signs of the disease but there is family history of the disease eg. haemophilia

allows people to make informed decisions regarding offspring. it gives information on the probability of having offspring with a disorder

80

prenatal screening and diagnosis

this is offered to women during her (...) could include (...) (...) and (...). these tests could indicate the (...) status of an embryo or fetes, where a specific genetic disorder is (...) or it (...)

make (...) whether to (...) with (...) or (...) if a condition is detected. this puts an (...) on prospective parents

prenatal screening and diagnosis

this is offered to women during her pregnancy, tests could include blood tests, ultrasound screening and cell free dna screening. these tests could indicate the genetic status of an embryo or fetes, where a specific genetic disorder is suspected or it signals a birth defect

make informed decisions whether to continue with pregnancy or abort if a condition is detected. this puts an emotional burden on prospective parents

81

predictive screening

to detect (...) associated with disorders where (...) that (...). often requested by people with (...) of a disorder eg. (...) is a dominant inherited disease with onset in middle age

source of (...) for people who test positive. (...) of results to (...) may (...) against a person. invasion of privacy

predictive screening

to detect mutations associated with disorders where symptoms that appear later in life. often requested by people with family history of a disorder eg. Huntington's disease is a dominant inherited disease with onset in middle age

source of anxiety and stress for people who test positive. misuse of results to third parties may discriminate against a person. invasion of privacy

82

embryo biopsy

or (...) is a tests that involves the (...) of a (...) from an (...) conceived through (...). the embryo is tested for genetic disorders before it is implanted into a woman uterus.

make (...). (...) embryo without disorders. (...)???

embryo biopsy

or preimplantation genetic diagnosis is a tests that involves the removal of a single cell from an embryo conceived through IVF. the embryo is tested for genetic disorders before it is implanted into a woman uterus.

make informed decisions. choose embryo without disorders. right to life???

83

dna profiling

a process used to compare the base sequence of two or more individuals

84

dna profiling
- (...) dna from (...) individuals based on (...) regions; (...) or (...)
- (...) of variation uses a combination of a (...), each (...) to a (...)

dna profiling
- identifies dna from different individuals based on variable regions; STR or microsatellites
- detection of variation uses a combination of a single locus probe, each specific to a different STR

85

- dna profiling makes use of (...) which are (...)
- dna fingerprinting makes use of (...), which are (...)

- dna profiling makes use of micro satellites which are 2-5bp
- dna fingerprinting makes use of mini satellites, which are 9-80bp

86

STR
definition?
- (...) greatly between individuals and each variation is a (...)
- the (...) at an (...) are named according to the (...)

hypervariable regions of chromosomes where sequences of jus two or 5 bp are repeated over and over- these are microsatellites

- vary greatly between individuals and each variation is a distinct allele
- the alleles at an STR locus are named according to the number of repeats

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DNA profiling:
- at each (...), an individual is either (...) or (...) and so can have a maximum of just (...) different alleles
- these alleles are inherited in a (...) fashion, and are (...); that is where (...) different alleles of the (...) are present, (...) can be (...)

DNA profiling:
- at each STR locus, an individual is either homozygous or heterozygous and so can have a maximum of just two different alleles
- these alleles are inherited in a mendelian fashion, and are codominant; that is where two different alleles of the same STR marker are present, both can be detected

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DNA profiling:
- DNA profile identifies (...) in every person's (...) and other organism. it is a kind of (...)

DNA profiling:
- DNA profile identifies natural variation in every person's DNA and other organism. it is a kind of bar code.

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DNA profiling: Source of dna
- dna used to create a profile cam come from either the (...) or the (...)
(...) contains (...) that can identify individuals with (...) because all individuals have a (...)
- (...) dna is not used to identify individual people, as(...) of the same (...) line will have the same (...)
- mtDNA is generally used to (...) or (...)

DNA profiling: Source of dna
- dna used to create a profile cam come from either the nucleus or the mitochondria
nuclear dna contains STRs that can identify individuals with accuracy because all individuals have a unique dna profile
- mitochondrial dna is not used to identify individual people, as family members of the same maternal line will have the same base sequences
- mtDNA is generally used to identify victims of mass disasters or badly decomposed bodies where the victims cannot be identified through physical means

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DNA profiling: uses of dna profiling
1. (...)-solving a crime
2.(...)- who's your daddy
3. (...)- identification of victims
4. (...)- missing persons or lost soldiers
5. (...)- ensure you have the prized animal
6. protecting Australia's wildlife-protect against unlawful collection and exportation
7. examination of ancient dna- answers historical questions

DNA profiling: uses of dna profiling
1. forensic investigations-solving a crime
2. paternity cases- who's your daddy
3. mass disasters- identification of victims
4. identification of human remains- missing persons or lost soldiers
5. pedigree analysis- ensure you have the prized animal
6. protecting Australia's wildlife-protect against unlawful collection and exportation
7. examination of ancient dna- answers historical questions

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DNA profiling for forensic investigations in australia:
- (...) from different chromosomes are used. these STRs were chosen because they are: (...), (...), (...), (...)(...)
- in addition, a tenth marker is used for (...) identification using the (...) gene X chromosome (...) bases and Y chromosome (...) bases

DNA profiling for forensic investigations in australia:
- nine STRs from different chromosomes are used. these STRs were chosen because they are: reproducible, robust, easy to score, highly informative, have a low mutation rate
- in addition, a tenth marker is used for gender identification using the Amel gene X chromosome 107 bases and Y chromosome 113 bases

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DNA profiling for forensic investigations in australia: The technique used

- each (...) is (...) by (...) using a specific(...)
- products of PCR undergo (...) and the(...)will emit green, blue or yellow colour when exposed to uv light
- a person shows one or two peaks where a peak corresponds to an allele depending on whether they are (...) or (...)
- for the Amel gender marker, if just a (...) peak with a size of (...) appears on the profile, the person is female; if two peaks are detected at (...) and the second at (...) then the person is male
- the dna profile is then (...) for the nine STR loci and the Amel gene

DNA profiling for forensic investigations in australia: The technique used

- each STR loci is amplified by PCR using a specific fluorescent primer
- products of PCR undergo electrophoresis and the fluorescent primers will emit green, blue or yellow colour when exposed to uv light
- a person shows one or two peaks where a peak corresponds to an allele depending on whether they are homozygous or heterozygous
- for the Amel gender marker, if just a single peak with a size of 107bp appears on the profile, the person is female; if two peaks are detected at 107 and the second at 113bp then the person is male
- the dna profile is then produced for the nine STR loci and the Amel gene

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DNA profiling for forensic investigations in australia: national DNA data banks

- many countries have set up national dna databanks which hold the dna profiles of people convicted of serous crimes and the dna profiles from scenes of crime
- this resource can be used as (...) to link people to scenes of crime, to link one crime scene to the scene of another crime, to clear innocent people and to assist in convicting the guilty
- in australia, the collection of dna samples is done in accordance with (...) and dna profiles are prepared using the (...)i at accredited forensic labs
- this info is digitally stored at the national criminal investigation DNA database (...) where it is stored (...) to safeguards ones privacy

DNA profiling for forensic investigations in australia: national DNA data banks

- many countries have set up national dna databanks which hold the dna profiles of people convicted of serous crimes and the dna profiles from scenes of crime
- this resource can be used as evidence to link people to scenes of crime, to link one crime scene to the scene of another crime, to clear innocent people and to assist in convicting the guilty
- in australia, the collection of dna samples is done in accordance with legislation and dna profiles are prepared using the agreed standard STR loci at accredited forensic labs
- this info is digitally stored at the national criminal investigation DNA database NCIDD where it is stored anonymously to safeguards ones privacy

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Issues with DNA profiling:
- debate exists whose dna profiles should remain in a national data bank, from individuals convicted of serious crimes to individuals who have been accused but not convicted
- concerns have been expressed that wider implementation of familial searching will cause investigators to rely too much on genetic leads at the expense of other potentially more useful lines of investigations
- the use of familiar searching also raises the issue of whether relatives of convicted criminals whose DNA is in databanks and who can be reached through familial searching are being unfairly discriminated against
- concerns have been raised about predictive testing of dna samples to identify aspects of a persons phenotype because phenotype is not determined exclusively by ones dna but is also affected by many environmental factors and this is still a science under development
- other issues raised relate to the possibility of reinforcing prejudices gains certain groups within the community and promoting racial stereotypes if DNA samples are used to determine genetic ancestry and whether this info is made public is made public in relation to individuals suspected of crime

Issues with DNA profiling:
- debate exists whose dna profiles should remain in a national data bank, from individuals convicted of serious crimes to individuals who have been accused but not convicted
- concerns have been expressed that wider implementation of familial searching will cause investigators to rely too much on genetic leads at the expense of other potentially more useful lines of investigations
- the use of familiar searching also raises the issue of whether relatives of convicted criminals whose DNA is in databanks and who can be reached through familial searching are being unfairly discriminated against
- concerns have been raised about predictive testing of dna samples to identify aspects of a persons phenotype because phenotype is not determined exclusively by ones dna but is also affected by many environmental factors and this is still a science under development
- other issues raised relate to the possibility of reinforcing prejudices gains certain groups within the community and promoting racial stereotypes if DNA samples are used to determine genetic ancestry and whether this info is made public is made public in relation to individuals suspected of crime

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dna profiling in animals: various purposes
- prosecuting people who are trading illegally in wildlife
- confirming the parentage pf valuable breeding stock. breeders pf pedigreed animals such as cattle and horses need to know the sire; father and dam; mother of the animals they breed
- in the case of a dog attack, matching saliva traces left on a victim to a particular dog
- domestic pets leave traces, such as hair, saliva, and blood, on clothing, carpets and furnishings. dna from these traces can provide forensic evidence to link human suspects to the scene of a crime or to a victim

dna profiling in animals: various purposes
- prosecuting people who are trading illegally in wildlife
- confirming the parentage pf valuable breeding stock. breeders pf pedigreed animals such as cattle and horses need to know the sire; father and dam; mother of the animals they breed
- in the case of a dog attack, matching saliva traces left on a victim to a particular dog
- domestic pets leave traces, such as hair, saliva, and blood, on clothing, carpets and furnishings. dna from these traces can provide forensic evidence to link human suspects to the scene of a crime or to a victim