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BIOL 2312 Spring 2023 > bio lab final > Flashcards

bio lab final Flashcards

1
Q

ph<pI

A

positively charged

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2
Q

ph>pI

A

negatively charged

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3
Q

how many times do you wash after seconary antibody

A

2

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4
Q

positive control

A

contains variable being tested for, demonstrates positive result, confirms everything is running correctly

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5
Q

negative control

A

does not contain tested variable, confirms no false positives, demonstrates negative results

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6
Q

Reusing a pipet tip when transferring controls and samples from tubes to wells

A

is more likely to cause false positives than false negatives.

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7
Q

Which of the following errors would result in a negative result for all samples in an ELISA (including positive controls)? Select all that apply.

A

Failure to add antigen.

Correct
Failure to add funtional primary antibody.

Correct
Failure to add functional secondary antibody.

Correct
Failure to add functional substrate.

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8
Q

Which of the following errors would result in positive results for all samples in an ELISA assay (including negative controls)?

A

Failure to perform the wash step after incubation with secondary antibody.

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9
Q

primase

A

makes first few nucleotides

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10
Q

steps of pcr

A

denaturing at 90-95, annealing at 55, extension at 72

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11
Q

how many genes in mt dna?

A

37, 13 protein, others for trna or rrna.

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12
Q

agarose concentration

A

0.5%>10 kb
1% for 500-10 kb
2% for smaller than 1 kb

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13
Q

chelex

A

removes metal ions that may catalyze dna digestion

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14
Q

mg2+ function

A

cofactor, facilitates nucleotide phosphate bonds between primer and incoming nucleotides

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15
Q

components of pcr rxn

A

dna, nucleotides, buffer with mg2+, primers, dna polymerase

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16
Q

steps for a pcr reaction

A

incubation with cacl2, cells, and the plasmid– cacl2 makes the cells competent
heat shock to stress cells and destabilize membranes
recovery– incubation for gene expression

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17
Q

arabinose function

A

binds to protein called arac on pbad so it allows binding of rna polymerase

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18
Q

griffith

A

found that combining smooth killed pneumococci with nonvirulent rough pneumococci caused death. transforming principle present

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19
Q

avery, macleod, mccarty

A

s-strain cells fractionated into classes of molecules and only dna transformed r strain into s strain

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20
Q

cells can be made permeable to dna using

A

calcium chloride, heat shock, electroporation, protoplast formation, microprojectiles

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21
Q

components of plasmid

A

origin of replication, gene of interest, genes with selective advantage, and polylinker area with multiple endonuclease cleavage areas

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22
Q

successful transformation needs

A

host cells, plasmid with selectable marker

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23
Q

constitutive gene

A

constantly expressed

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24
Q

inducible

A

gene only in certain conditions

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25
Q

total amount of pglo used

A

volume*concentration

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26
Q

fraction of dna

A

volume spread/ total volume (cacl2+lb broth+plasmid dna)

27
Q

micrograms spread

A

amount of dna*fraction

28
Q

transformation efficiency

A

colonies/micrograms spread

29
Q

plasmid pglo characteristics

A

Plasmid: pGLO - selectable marker (bla -
ampicillin resistance gene), gene for GFP,
origin of replication (ori), polylinker area,
special gene regulation system (Promotor -
P(BAD) and araC gene)

30
Q

beer’s law

A

a=bec (b=1 cm pathlength, c-concentration)

31
Q

kmno4 and chemicals

A

plastic cuvettes

32
Q

proteins

A

quartz cuvettes and plate readers

33
Q

nucleic acids

A

low volume spectrophotometer

34
Q

ph of acid phosphatase reaction

A

4.5
nitrophenyl phosphate to nitrophenol and phosphate

35
Q

acidic amino acids

A

aspartic acid, glutamic acid

36
Q

basic amino asids

A

lysine, arginine, histidine

37
Q

At what temperature is Taq DNA polymerase most active?

A

72-75

38
Q

The two essential components that need to be added in addition to the “Dream Taq Green PCR Master Mix” to set up the PCR reaction are:

A

Primers and DNA template

39
Q

The two primers for one PCR reaction should be complementary to each other.

A

F

40
Q

The direction of synthesis of the new DNA strand in each cycle of the PCR is from 5’ to 3’ end.

A

T

41
Q

Which of the following are true in regards to mitochondrial DNA (mtDNA)? Choose all that apply.

A

The mitochondrial genome contains 37 genes.

Correct
There are more copies of mt genes than nuclear genes per cell.

42
Q

Makes cells permeable to plasmid DNA

Correct C.
CaCl2

Correct C.
CaCl2

Selects for the growth of transformed cells

Correct E.
Ampicillin

Correct E.
Ampicillin

Provides the GFP gene

Correct B.
pGLO plasmid

Correct B.
pGLO plasmid

Induces GFP gene expression

Correct A.
Arabinose

Correct A.
Arabinose

A
43
Q

To increase the efficiency of transformation, cells will be placed in a 42°C water bath for 50 seconds during which step of the procedure?

A

heat shock

44
Q

Which of the following is a method for making cells competent (permeable to plasmid DNA) ?

A

electroporation, microprojectile, protoplast formation

45
Q

In our experiment, expression of the beta-lactamase gene (from the plasmid) is constitutive.

A

T

46
Q

HB101 is a strain of E. coli that is naturally antibiotic resistant.

A

F

47
Q

During a transformation experiment with pGLO, the gene for GFP will remain turned off if arabinose is present in the media.

A

F

48
Q

Which of the following wavelength ranges describes visible light?

A

400-700

49
Q

x axis absorption and standard curve

A

wavelength, concentration

50
Q

We will discover the lambda max (λmax ) of a chemical solution by generating and analyzing the absorption spectrum.

A

t– accuracy and sensitivity

51
Q

The molar absorptivity of a chemical (ε) is always constant at any wavelength (ie – it would be the same at 300nm and 400nm and 500nm)

A

F

52
Q

Autozero (pressing Measure Blank) is required when a new wavelength is set

A

T

53
Q

The initial velocity of an enzyme catalyzed reaction is determined by

A

calculating the amount of product formed over time while the amount of product is increasing in a linear manner

54
Q

Which of the following influence the rates of enzyme-catalyzed reactions?

A

:
Correct
Concentration of substrate

Correct
Presence of activators or inhibitors

Correct
Temperature and pH

Correct
Amount of enzyme

55
Q

The experiment you will be performing in lab assays the reaction rate of wheat germ acid phosphatase at a temperature of [a]°C.

A

37

56
Q

Question Correct Match Selected Match
Nitrophenyl phosphate

Correct C.
substrate

Correct C.
substrate

Nitrophenol

Correct A.
product

Correct A.
product

Acid phosphatase

Correct D.
enzyme

Correct D.
enzyme

1.5% KOH

Correct B.
stops reaction and allows absorbance of product to be measured

Correct B.
stops reaction and allows absorbance of product to be measured

A
57
Q

nitrophenol standard curve

A

x=cocnentration
y=absorbance

58
Q

The isoelectric point (pI) of an amino acid is defined as the pH at which the amino acid does not migrate in an electric field or the pH at which an amino acid bears no net charge.

A

T

59
Q

Which of the following four proteins in Part A of today’s experiment is the most acidic protein?

A

bovine albumin

60
Q

1 out of 1 points

Correct
If amino acid Lysine with a pI of 9.7 is dissolved in a gel running buffer of pH 6; at this pH, Lysine is a___________ and it will migrate toward____________ during the native agarose gel electrophoresis

A

cation, cathode

61
Q

If your group is assigned to prepare 60 ml of 1.2% agarose gel solution for part B, what materials are required to prepare the gel solution. Select the amount of agarose and the amount of the correct buffer needed.

A

Correct
0.72 g of agarose

Correct
60 ml of electrophoresis buffer with pH at 9.2

62
Q

Which two of the following statements about the separation of molecules are true?

A

Separation of Protein using “Native” or “non-denaturing” gel electrophoresis is on the basis of the net charges

Correct
Separation of DNA molecules using gel electrophoresis is on the basis of sizes

63
Q

What is the basis of separation of the HbA and HbS in Part B of today’s experiment?

A

charge

64
Q

Incubation: the cells mixed with a transformation solution of CaCl2 (calcium chloride) and the plasmid.
* Heatshock: the cells and DNA mixture are then subjected to a treatment at 42ºC for 50 seconds; the temperature changes destabilize cell membranes and create thermal imbalance.
* Recovery: a short incubation period to begin expressing their newly acquired genes

A

BIOL 2312 Spring 2023 (3 decks)

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