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BMME 485 > Exam 1- Lecture 4 > Flashcards

Exam 1- Lecture 4 Flashcards

1
Q

What acid is used to separate DNA and RNA

A

Phenol

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2
Q

How is phenol used to separate DNA and proteins

A

phenol is mixed with aqueous sample of DNA and proteins
proteins dissolve in phenol layer
nucleic acids form aqueous layer
layers separated through centrifugation

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3
Q

After using phenol to separate DNA from proteins, what enzyme is used to digest RNA into ribonucleotides

A

RNase

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4
Q

To separate DNA from ribonucleotides, alcohol is added and

A

DNA falls out of aqueous phase and is isolated
ribonucleotides stay soluble in solution

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5
Q

What is used to separate DNA fragments by size

A

gel electrophoresis

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6
Q

How does gel electrophoresis separate DNA fragments by size

A

electric current is passed through gel, negatively charged fragments move away from negative electrode

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7
Q

Do shorter DNA molecules move farther or closer to

A

they move farther than longer molecules

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8
Q

What are used to cut bacterial enzymes at specific recognition sites on DNA

A

restriction enzymes/endonucleases

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9
Q

What type of DNA can restriction enzymes not cut

A

their own DNA because it is methylated

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10
Q

What type of restriction enzyme is most useful for genetic engineering

A

Type 2

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11
Q

Where do type 2 restriction enzymes cut DNA

A

in the middle of recognition sequence

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12
Q

What is the function of DNA ligase

A

to link or ligate two different DNA fragments that have been cut using restriction enzymes

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13
Q

How could you make ligase more efficient

A

By having the two DNA fragments have the same overhanging sticky ends

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14
Q

In order to detect specific DNA molecules radioactively what is inserted into the molecules during replication

A

phosphate group
phosphorothioate group

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15
Q

What does autoradiography identify radioactive DNA as

A

“hot”

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16
Q

What will radioactive DNA look like during autoradiography?

A

It will turn gel surrounding it black

17
Q

How is fluorescence used to determine nucleic acids

A

fluorescently tagged nucleotides incorporated into 3’ end of DNA
beam of light excites fluorescent tag
releases light

18
Q

What is a Southern blot used for

A

form hybrid DNA molecule to determine if sample DNA has homologous sequence to another DNA molecule

19
Q

What are Northern blots used for?

A

determine if sample of mRNA has homologous sequence to DNA probe

20
Q

Why are Northern blots preferred to use for larger genomes

A

Because all of the introns are removed

21
Q

What is the FISH technique

A

labeled probe is incubated with cells that have had their DNA denatured by heat
probe hybridizes to its homologous sequence on chromosome

22
Q

What are cloning vectors

A

specialized plasmids that will hold any piece of foreign DNA for further study or manipulation

23
Q

What is the polylinker region

A

region in cloning vector with multiple restriction enzyme sites

24
Q

What are two ways inserts are detected within plasmids

A

resistance to antibiotic
synthesis of B-galactosidase

25
Q

What happens during the resistance to antibiotic for insertional activation?

A

cells carrying insert are only resistant to first antibiotic not the second

26
Q

What happens during the synthesis of B-galactosidase for alpha complementation?

A

in cells with insert, B-galactosidase is inactive, cells remain white

27
Q

YAC stands for

A

yeast artificial chromosomes

28
Q

What are the two types of YAC

A

circular form for growing in bacteria
linear form for growing in yeast

29
Q

What type of artificial chromosomes can hold the largest amount of DNA

A

YACs

30
Q

How are BACs made

A

cut DNA of interest
linearize vector and mix with DNA of interest
transform mixture into bacteria
grow and isolate large amounts of DNA

31
Q

What is cDNA

A

copy of messenger RNA
no introns present

32
Q

What are the steps to make cDNA expression libraries

A

reverse transcriptase plus primers with oligo(dT) are added
oligo(dT) hybridizes to polyA tail and acts as primer for reverse transcriptase which makes cDNA
mRNA/cDNA heteroduplex is formed
mRNA strand removed
DNA polymerase I used to make opposite strand

33
Q

What is genome wide association study

A

study of genome-wide set of genetic variants in different individuals to see if there is any variant associated with trait

34
Q
A

BMME 485 (15 decks)

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