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Genetics Class > Lab Quiz 2 > Flashcards

Lab Quiz 2 Flashcards

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Biology 101 flashcards
1
Q

What are autotrophs

A

Bacteria have mutations that prevent them from synthesizing compounds needed for cell growth

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2
Q

What is a reverse mutation?

A

Can restore the ability of the bacterium to synthesize the essential compound

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3
Q

What is a prototroph

A

A bacteria which can grow without any nutrient supplementation

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4
Q

What are spontaneous mutations

A

Mutation events occurring in the absence of a mutagen. Can cause a lost gene to reappear

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5
Q

What is a negative control

A

A plate without the use of a mutagen. The average number of colonies on these control plates measure the rate of spontaneous mutation

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6
Q

What are positive control plates

A

Include the mutagen which are known to produce revertant colonies in the mutant bacterial strain

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7
Q

What is methyl methane sulfonate

A

Used for positive control, known as a mutagen

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8
Q

What was DMSO used for

A

Negative control, was used as a solvent for student sample. Needs to be negative control in order to ensure this isnt causing the mutations

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9
Q

What is the sterility control

A

Asses the sterility of each student substance

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10
Q

How do we know if a substance is toxic?

A

It requires a zone of inhibition

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11
Q

How do we know if a substance is mutagenic?

A

Has to be double number of colonies from negative control

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12
Q

What are the two types of mutagens?

A

Direct acting mutagens react directly with DNA to produce base changes

Indirect-acting mutagens are not themselves mutagens, but are converted during metabolism in the liver to chemicals that are mutagens

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13
Q

What does sodium nitrite do?

A

Causes deamination of cytosine to produce uracil

This introduces a base pair substitution mutation

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14
Q

What is acridine orange

A
  • Sandwiches itself between adjacent bases
  • Distorts the DNA helix and causes insertions and deletion during replication
  • Introduces a frameshift mutation
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15
Q

What were the three bacteria that we tested?

A
  • E coli (trp-)
    • Detects base pair
  • S. Typhimurium A (his-)
    • detects base pair
  • S Typhimurium B (his-)
    • Detect frameshift
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16
Q

Why did we use bacteria that could not synthesize certain amino acids?

A

Only bacteria that have undergone a reverse mutation will be able to synthesize the amino acid and grown on minimal medium

That is why we used trace amounts of amino acid, it is insufficient amount to allow auxotrophic bacteria to grow into colonies

17
Q

Trick for pedigree

A
18
Q

What are plasmids

A

non chromosomal supercoiled DNA

19
Q

What are recombinant plasmids

A

replication of the recombinant plasmid in bacteria cells provides a means to produce many copies or cline the DNA of interest

20
Q

What does TE buffer do?

A

eliminates proteins and other molecules found in the growth medium

21
Q

What is SDS for in plasmid extraction for

A

(detergent) Lyses cells by disrupting lipids in the cell membrane

22
Q

What is NaOH in plasmid extraction for

A

Denature proteins and DNA

23
Q

What is potassium acetate for?

A

used to neutralize the base and potassium reacts with SDS to form an insoluble precipitate that traps proteins, chromosomal DNA, and membrane but not plasmid or RNA

24
Q

What does low temperature do in plasmid isolation?

A

encourages precipitation

25
Q

During the first centrifugation, what do you keep?

A

Separates proteins, chromosomal DNA and cell membrane (pellet) from plasmid DNA and RNA (supernatant)

26
Q

What does use 95% ethyl alcohol do?

A

Precipitates plasmid DNA and RNA

27
Q

What does the second centrifugation do?

A

Separates plasmid DNA and RNA (pellet_ from ethyl alcohol and inorganic salt (supernatant). Keep pellet

28
Q

Why do we dry?

A

Ethyl alcohol will interfere with enzymatic digestion

29
Q

What is RNase for?

A

Used to destroy RNA

30
Q

What are the steps for DNA plasmid extraction

A
31
Q

What is a restriction endonuclease?

A

An enzyme that makes a double-stranded cut within an intact DNA double helix at a specific nucleotide sequence called restriction site.

32
Q

What is BamH1 do?

A

recognizes the six base pair palindromic sequence

  • GGATCC
  • CCTAGG
  • G GATCC
  • CCTAG G
33
Q

What determines how rapidly DNA moves through gel matrix

A

Size/weight/shape of the DNA

Circular shape will migrate faster than linear

Genetics Class (10 decks)

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