bio lab final

Question 1

ph<pI

Answer 1

positively charged

Question 2

ph>pI

Answer 2

negatively charged

Question 3

how many times do you wash after seconary antibody

Answer 3

2

Question 4

positive control

Answer 4

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

Question 5

negative control

Answer 5

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

Question 6

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

Answer 6

is more likely to cause false positives than false negatives.

Question 7

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

Answer 7

Failure to add antigen.

Correct
Failure to add funtional primary antibody.

Correct
Failure to add functional secondary antibody.

Correct
Failure to add functional substrate.

Question 8

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

Answer 8

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

Question 9

primase

Answer 9

makes first few nucleotides

Question 10

steps of pcr

Answer 10

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

Question 11

how many genes in mt dna?

Answer 11

37, 13 protein, others for trna or rrna.

Question 12

agarose concentration

Answer 12

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

Question 13

chelex

Answer 13

removes metal ions that may catalyze dna digestion

Question 14

mg2+ function

Answer 14

cofactor, facilitates nucleotide phosphate bonds between primer and incoming nucleotides

Question 15

components of pcr rxn

Answer 15

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

Question 16

steps for a pcr reaction

Answer 16

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

Question 17

arabinose function

Answer 17

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

Question 18

griffith

Answer 18

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

Question 19

avery, macleod, mccarty

Answer 19

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

Question 20

cells can be made permeable to dna using

Answer 20

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

Question 21

components of plasmid

Answer 21

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

Question 22

successful transformation needs

Answer 22

host cells, plasmid with selectable marker

Question 23

constitutive gene

Answer 23

constantly expressed

Question 24

inducible

Answer 24

gene only in certain conditions

Question 25

total amount of pglo used

Answer 25

volume*concentration

Question 26

fraction of dna

Answer 26

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

Question 27

micrograms spread

Answer 27

amount of dna*fraction

Question 28

transformation efficiency

Answer 28

colonies/micrograms spread

Question 29

plasmid pglo characteristics

Answer 29

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)

Question 30

beer's law

Answer 30

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

Question 31

kmno4 and chemicals

Answer 31

plastic cuvettes

Question 32

proteins

Answer 32

quartz cuvettes and plate readers

Question 33

nucleic acids

Answer 33

low volume spectrophotometer

Question 34

ph of acid phosphatase reaction

Answer 34

4.5 nitrophenyl phosphate to nitrophenol and phosphate

Question 35

acidic amino acids

Answer 35

aspartic acid, glutamic acid

Question 36

basic amino asids

Answer 36

lysine, arginine, histidine

Question 37

At what temperature is Taq DNA polymerase most active?

Answer 37

72-75

Question 38

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:

Answer 38

Primers and DNA template

Question 39

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

Answer 39

F

Question 40

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

Answer 40

T

Question 41

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

Answer 41

The mitochondrial genome contains 37 genes. Correct There are more copies of mt genes than nuclear genes per cell.

Question 42

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

Question 43

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?

Answer 43

heat shock

Question 44

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

Answer 44

electroporation, microprojectile, protoplast formation

Question 45

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

Answer 45

T

Question 46

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

Answer 46

F

Question 47

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

Answer 47

F

Question 48

Which of the following wavelength ranges describes visible light?

Answer 48

400-700

Question 49

x axis absorption and standard curve

Answer 49

wavelength, concentration

Question 50

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

Answer 50

t-- accuracy and sensitivity

Question 51

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)

Answer 51

F

Question 52

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

Answer 52

T

Question 53

The initial velocity of an enzyme catalyzed reaction is determined by

Answer 53

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

Question 54

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

Answer 54

: Correct Concentration of substrate Correct Presence of activators or inhibitors Correct Temperature and pH Correct Amount of enzyme

Question 55

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

Answer 55

37

Question 56

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

Question 57

nitrophenol standard curve

Answer 57

x=cocnentration y=absorbance

Question 58

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.

Answer 58

T

Question 59

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

Answer 59

bovine albumin

Question 60

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

Answer 60

cation, cathode

Question 61

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.

Answer 61

Correct 0.72 g of agarose Correct 60 ml of electrophoresis buffer with pH at 9.2

Question 62

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

Answer 62

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

Question 63

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

Answer 63

charge

Question 64

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