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Biochemistry Lab > FINAL review > Flashcards

FINAL review Flashcards

(62 cards)

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1
Q

what is purpose of Lowry assay

A

determine protein level in solution

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2
Q

What color is generated in Lowry assay and what absorbance level is it measured

A

blue color; 750nm and 540 nm

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3
Q

the total protein concentration in Lowry assay is exhibited by what

A

color change of sample solution in proportion to protein concentration

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4
Q

how is absorbance measured

A

using spectrophotometer

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5
Q

Lowry reagent \/ Colin phenol is

A

a strong acid

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6
Q

in bradford assay, the absorbance is at

A

595 nm

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7
Q

what amino acids are required for Lowry assay

A

tyrosine and tryptophan

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8
Q

Lowry assay requires samples containing only

A

10-200 ug/ml of protein

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9
Q

Lowry assay generates ____ color; biuret assay results in _____ color

A

blue; purple

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10
Q

what two groups are found in an amino acid

A

amino group and carboxylic acid group

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11
Q

titration curve is a plot of what

A

pH of weak acid against degree of neutralization of acid by strong base

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12
Q

what does a zwitterion contain

A

a protonated amino group and deprotonated carboxyl group

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13
Q

what titrants are used in amino acid titration

A

1M KOH

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14
Q

how is unknown amino acid identified in amino acid titration

A

calculating its molecular mass

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15
Q

ninhydrin reaction is what type of reaction

A

redox reaction

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16
Q

what groups are involved in ninhydrin reaction

A

amino group of sample and ninhydrin reagent

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17
Q

what is end product formed similar to

A

di-dehydrin

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18
Q

di-ketohydrin has what color and what is it also known as

A

deep blue pigmentation; ruhemann’s purple

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19
Q

what is ninhydrin reagent used for

A

qualitative and quantitive colorimetric analysis for amino acids and sometimes for peptides and proteins

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20
Q

what molecules are used in part A of ninhydrin reaction

A

glucose, serine, and glycine

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21
Q

what technique is used and what is its purpose in ninhydrin reaction

A

thin layer chromatography; used to separate and identify amino acids

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22
Q

what result is a positive test in ninhydrin reaction

A

purple color

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23
Q

what is function of protease

A

enzyme that catalyzes proteolysis or breakdown of protein (break down peptide bonds)

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24
Q

how is protease activity measured

A

colormetric method

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25
what are protease enzymes involved in
cell division, growth, apoptosis, and migration
26
TCA is used in protease activity to do what
stop the reaction by precipitating enzymes and unreacted substrates
27
ninhydrin reagents can be used in protease activity for what
quantification of TCA soluble peptides
28
what enzymes are used in protease activity
trypsin and substrate casein
29
enzyme kinetics is study of
rates of enzyme catalyzed chemical reactions
30
trypsin plays role of ___ and casein plays role of _____
enzyme; substrate
31
what molecule is used to stop enzyme catalyzed reaction
trichloroacetic acid
32
molecular weight of casein is
33,300 daltons
33
what is name of v vs [s] plot
Michaeles Menten plot
34
velocity is ____ variable and substrate conc. is ___ varible
dependent; independent
35
galactose is _____ because its aldehyde group is easily oxidized to carboxylic acid
reducing sugar
36
what result does sucrose give in resorcinol test
positive result
37
sucrose and trehalose are what type of sugars
non-reducing sugars
38
purpose of carbohydrate analysis experiment
test characteristics of sugars
39
Benedict's reagents tests is for
reducing sugar
40
what is positive result in beneditcs reagent; Molisch test; resorcinol test; iodine-KI test
greenish red color (reducing sugar); red violet color (carbohydrate presence); red for ketone sugar and identifies between aldose and ketone sugar; blue color in presence of amylose/starch
41
what is purpose of DNA plasmids isolation and PCR
extract DNA plasmids from cell cultures and measure DNA concentration
42
what are steps for PCR
denaturation, annealing and extenstion
43
SDS has what purpose in DNA isolation lab
detergent that destabilizes cell wall and cause cell wall to burst and release concents
44
TE contains what
Tris-HCl which is a buffer to obtain correct pH
45
EDTA is what
chelates ions needed by DNAse, inactivate DNAses and keep DNA from degrading
46
another name for DNA fragment is
amplicon
47
DNA absorbs light at
260 nm and protein at 280nm
48
Agarose gel electrophoresis is good at separating fragments ranging from
100 bp to 25kb
49
purpose of DNA agarose gel lab
check PCR product and DNA size
50
what is DNA agarose gel made of
0.5 g agarose, TBE buffer, and ethidium bromide
51
what is purpose of using UV light in DNA agarose gel
visualize DNA
52
ethidium bromide bonded to DNA will glow what color at what wavelength
orange; 302
53
comparing mobility of DNA of known size to mobility of
your PCR fragment
54
purpose of gel filtration chromatography is
purification of biological macromolecules
55
gel filtration chromatography can separate protein of
different masses
56
how is molecular mass of unknown protein found
by comparing to elution volumes of molcular mass standards
57
how is hydrodynamic radius determined
by fluidity instruments ...
58
AGE stands for
polyacrylamide gel electrophoresis
59
the unique migration rate of protein in SDS PAGE is found by
size of protein
60
SDS is considered what
an anionic deteregent
61
large negative charge of SDS can
impart once bounded to protein
62
what is SDS PAGE used for
separate molecules of DNA RNA or protein on basis of size or electric hcarge

Biochemistry Lab (1 decks)

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