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bio 243 > Lab final > Flashcards

Lab final Flashcards

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Biology 101 flashcards
1
Q

what are the two thiings dtetergent due in DNA extraction?

A
  1. lyses the DNA (breaks down cell membrane)
  2. Denautures histones and seperates them in the DNA
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2
Q

what does isopropanol do to the extracted DNA?

A

makes DNA precipitate out of the extraction buffer

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3
Q

what does the 70% ethanol do to the extracted DNA?

A

washes off proteins, RNA, and lipids

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4
Q

what does the addition of NaOH do to affect the florescence of the DNA?

A

NaOH denautures the DNA so that double stranded DNA can;t interact with the dye.

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5
Q

How does the NaOh denauture the DNA?

A

NaOH removes protons from nitogenous bases of DNA because it is a strong base. This disrupts the hydrogen bonding necessary for double stranded stabilization

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6
Q

which plate do you expect to find bacteria most like the wildtype bacteria?

A

LB/-pGLO plate

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7
Q

why would the LB/-pGLO plate have the most wildtype bacteria?

A

DNA plasmid was not added to the -pGLO therefore expression of the gene encoding GFP is not present.

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8
Q

which plate would most likely have genetically transformed bacterial cells?

A

both LB/amp and LB/amp/ara plates

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9
Q

Why would the +pGLO plates have genetically transformed bacteria cells?

A

Because they both allow for bacterial growth
-LB/amp expresses an ampicilin resistance gene enabling growth from the resistance
-LB/amp/ara allows GFP gene transcription when arabinose is present

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10
Q

which plates should be compared to determine if any genetic transformation has occured?

A

LB/amp/-pGLO and LB/amp/+pGLO

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11
Q

why would you compare LB/amp/-pGLO and LB/amp/+pGLO to see if genetic transformation has occured?

A

the -pGLO plate wont express ampicilin gene and will not grow while the +pGLO plate will have resistance to ampicilin and will grow GFP

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12
Q

what is a control plate?

A

a plate that serves purpose as to compare the outcome of the experiment to a baseline and help interperet results

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13
Q

what is the control plate in this experiement?

A

LB/-pGLO

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14
Q

why is the LB/-pGLO the control plate?

A

because plasmid is needed for growth and this plate is not manipulated by amp or plasmid which means no change will occur

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15
Q

How is pGLO expression in the transformed bacteria being induced?

A

the araC is added to the ara plate which allows GFP gene transcription, followed by production of GFP= bright green color under blue light

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16
Q

what is the purpose of the CaCl2 transformation solution?

A

it promotes plasmid DNA to bind to surface of bacteria cells by neutralizing (-) charge

17
Q

what is the purpose of the heat shock to the DNA?

A

it facilitates entry of DNA into bacterial cells, it makes membranes porous and changes fluitidy of lipids

18
Q

what bacterial structures must the pGLO plasmids pass through in order to enter the bacterial cells?

A

the cell membrane and cell wall

19
Q

what two things were done to help the plasmid enter the cell?

A
  1. added transformation solution with CaCl2
  2. heat shocked
20
Q

why was the microcentrifuge tube incubated for 10 minutes before adding to agar plates?

A

to allow the cells time to produce beta-lactamase

21
Q

would you expect to observe wildtype bacteria on plates that didn’t contain ampicillin or arabinose?

A

yes, LB/-pGLO would have wildtype bacteria because DNA of the plasmid was not expressed (nothing to make it change)

22
Q

what information would the results on the LB/amp/-pGLO plate provide?

A

it showed no growth meaning the introduction of antibiotic ampicillin could not be broken down and there was no plasmid on the plate

23
Q

what information would the results on the LB/amp/+pGLO plate provide?

A

bacteria can grow because plasmid was present and it was able to create a resistence to ampicilin and genes for GFP are expressed

24
Q

would there be any major difference in the number of coloonies between the LB/amp/+pGLO plate and the LB/amp/ara/+pGLO plate?

A

no there would be no difference

25
Q

would there be a difference in the phenotype of the bacteria on the two +pGLO plates?

A

-the LB/amp/+pGLO wouldn’t glow because antibiotic amp inhinits a gene expression that causes a glow
-the LB/ara/amp/+pGLO does glow because it has sugar arabinose that allows the transcription of the GFP gene

26
Q

what does NaCl do to DNA?

A

forms favorable electrostatic interactions with DNA and histones to seperate histones from DNA

27
Q

why use strawberries for DNA extraction?

A

have multiple copies of each chromosome
polyploidy
gives us more dna to extract

28
Q

what does dye do?

A

gives off bright yellow-gold when bound to DNA and exposed to blue light

29
Q

how does the dye form light?

A

interacts with (-) backbone of DNA and tightly stacked bases and only with double stranded DNA

30
Q

what is aseptic technique?

A

a technique that ensures you don’t contaminate yourself or your cultures

31
Q

what are the steps to aseptic technique?

A
  1. clean lab coat and fully buttoned
  2. no legs exposed. no open toed footwear, long hair tied back
  3. bench must be clear of non-essential items
  4. disinfect bench
  5. wash hands before and after lab
32
Q

what are the 3 different types of point mutations?

A

frameshift (addition and deletion)
stop
silent

33
Q

what is mitosis?

A

divides genetic material to daughter cells equally, daughter cells have the same genetic material as mother

34
Q

what is meiosis?

A

divides the genetic material among 4 daughter cells, each contains half the genetic material of mother

35
Q

why root tips?

A

tips of plant roots are rapidly growing tissues where dna replication mitosis and cell division occur at a high rate

36
Q

what is amp?

A

an antibiotic (combat bacterial growth)

37
Q

what is the trigger for bacterial growth?

A

arabinose

bio 243 (10 decks)

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