ch. 19 Genetic Technologies Flashcards Preview

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Flashcards in ch. 19 Genetic Technologies Deck (26)
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1
Q

what is used when many copies of DN are required for testing?

A

polymerase chain reaction (PCR)

2
Q

what does PCR require?

A

target DNA, lab-made primers which are complementary to the target sequence, nucleotides, Taq polymerase from Thermus aquaticus

3
Q

what is Taq?

A

thermophila, found in hot springs, function best at a very high temp

4
Q

what does heat do?

A

separates strands of target DNA, primers attach to target sequence, Taq polymerase adds nucleotides to target

5
Q

what is exponential amplification?

A

doubling with each cycle, usually requires at least 20 cycles

6
Q

What if you have RNA, but not DNA?

A

need to do reverse transcription to go from RNA to DNA; makes a compliementary DNA strand (cDNA) which then can be used for amplification of the gene

7
Q

what is southern blotting?

A

used to detect particular gene within DNA, provides info about # of copies of type of gene, identify homologous genes (sim but not identical), denture DNA run through gel electrophoresis transfer to membrane with a radiolabeled probe

8
Q

what charge does DNA have?

A

negative so top of gel negative and bottom positive so DNA drawn to gel

9
Q

What if southern blotting shows that the gene is there?

A

it may or may not show up on the northern blot. the protein may or may not show up on the western blot

10
Q

what is the western blot?

A

need to make an antibody that will recognize the protein a secondary antibody will recognize primary antibody and has dye on it.

11
Q

what is the last step of a western blot?

A

add aklaline phosphatase which pulls off of secondary antibody, which creates a color change (usually gray or black)

12
Q

t/f secondary antibodies are much less specific, bind to almost any primary antibody, so cheaper to make

A

true

13
Q

t/f restriction enzymes bind DNA at specific points and looks for symmetrical strands?

A

false- breaks

14
Q

In DNA sequencing what is dideoxyribonucleotides (ddNTPs)?

A

missing hydroxyl group from the 3’ carbon, ddATP, ddTTP, ddCTP, ddGTP

15
Q

what happens if there is no hydroxyl group?

A

polymerase will stop causing termination of growing chain of DNA.

16
Q

t/f in DNA sequencing of ddNTP each type of nucleotide is labeled with a specific color of fluorescence?

A

true

17
Q

which strands move to bottom of gel in test tube and detect fluorescence of each strand?

A

shorter

18
Q

what type of DNA is included from an organism of a different spieces?

A

recombinant DNA

19
Q

t/f Recombinant DNA has restriction enzymes that cut DNA at specific locations

A

true

20
Q

donor DNA complementary to “sticky” ends of host DNA are added and then joined by what in recombinant DNA?

A

ligase

21
Q

what is the last step of recombinant DNA?

A

divide to produce protein of interest

22
Q

t/f donor DNA is the gene you want to add to the cytoplasm and fits perfectly into the parts that are cut out from the cytoplasm?

A

false- plasmids

23
Q

what is an animal model?

A

insert genes into animals to produce a human disease such as Alzheimers

24
Q

what is a knockout in animal models?

A

both copies of the gene are replaced with inactivated mutated gene which tells us what a gene does

25
Q

what are some benefits of animal models?

A

easier than doing it on a human, much faster gene replication than in humans, life span is shorter (shorter timeframe to become mature), good sample size, larger animals have more similar DNA which gives better idea of how gene would affect human

26
Q

what are some drawbacks of animal models?

A

may not be able to get from a mouse exactly what would happen in humans