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Mol Bio Lab > (M) L2.1 : Gel Electrophoresis > Flashcards

(M) L2.1 : Gel Electrophoresis Flashcards

1
Q

commonly used to separate DNA fragments from each other

A

electrophoresis

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2
Q

we use electrophoresis to see the presence and sizes of ______

A

DNA fragments

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3
Q

can be used to analyze restriction-digestion of products

A

electrophoresis

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4
Q

Electrophoresis is a method of separating __________ _________ substances in a mixture

A

electrically charged substances

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5
Q

enumerate the three electrically charged substances mentioned

A

DNA, RNA, and proteins

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6
Q

where is the sample of the mixture is placed on?

A

supporting medium

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7
Q

use this card to familiarize examples of supporting medium (or enumerate if you’re a masochist)

A

paper
starch gel
cellulose acetate
agarose gel
polyacrylamide gel

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8
Q

Selecting the type of supporting medium depends on the _____

A

experiment

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9
Q

a technique used for separation of charged molecules such as DNA, RNA, or protein molecules through liquid or gel medium according to their size and charge using an electrical current

A

electrophoresis

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10
Q

negative or positive electrode?

net positive charge migrates toward the cathode?

A

negative electrode

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11
Q

negative or positive electrode?

net negative charge move toward the anode

A

positive electrode

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12
Q

this allows the biomolecules to migrate towards the electrode of opposite charge

A

electrostatic attraction

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13
Q

the ________ in electrical field depends on its net charge, size, shape, strength of electrical field, properties of supporting medium and temperature

A

rate of migration of an ion

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14
Q

factors that affect the rate of migration of an ion

A

net charge
size
shape
strength of electrical field

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15
Q

these molecules have positive or negative charges

A

biological molecules

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16
Q

what is the charge of biological molecules in acidic conditions?

A

positively charged

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17
Q

what is the charge of biological molecules in alkaline conditions?

A

negatively charged

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18
Q

who pioneered the work on moving boundary electrophoresis in 1930?

A

Arne Tiselius

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19
Q

What did Arne Tiselius developed after moving boundary electrophoresis?

A

zone method for the purification of biomolecules

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20
Q

when was tools for DNA gel electrophoresis was developed?

A

1970

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21
Q

who developed the tools for DNA gel electrophoresis in 1970

(3 people)

A

Phil Sharp
Joe Sambook
Bill Sudgen

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22
Q

what did the inventors of the tool for DNA gel electrophoresis use?

A

agarose gel from seaweed

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23
Q

Why did Arne Tiselius develop electrophoresis?

A

to study serum proteins

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24
Q

what are the 3 parts of electrophoresis?

A

voltage
supporting medium
buffer system

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25
this component of gel electrophoresis contains ions to conduct electricity
running buffer
26
type of electrophoresis in which supporting medium is a gel
gel electroforesis
27
the gel material acts as a __________ by retarding or completely obstructing the movement of macromolecules while allowing the smaller molecules to migrate freely
molecular sieve
28
T or F gel electrophoresis is widely used to combine proteins and nucleic acids
F (used to separate)
29
what are the 2 set-ups of gel electrophoresis
vertical and horizontal gel electrophoresis
30
type of gel used for nucleic acids
agarose
31
type of gel used for smaller nucleic acid
polyacrylamide
32
gel structure held together by covalent cross-linking of acrylamide and N,N'-methylene bisacrylamide
polyacrylamide gel (PAG)
33
T or F PAG can be used to separate DNA smaller than 1000 bp
F (less than 100 bp only)
34
component of discontinuous gel electrophoresis that contains a low percentage of acrylamide concentration and low pH
stocking gel
35
component of discontinuous gel electrophoresis that has a varying concentration of acrylamide and a higher pH
running / separating gel
36
T or F in PAG electrophoresis DNA mobility observed in the gel (rate of migration) are dependent on the electrical field of strength used for the electrophoresis
F (it's INDEPENDENT on the electrical field strength)
37
PAG electrophoresis depends on what? 3
size of analyte pore size concentration of acrylamide gel
38
T or F concentration of gel is inversely proportional to the pore size
T
39
what should be the concentration and pore size needed for analysis of high molecular weight
low gel concentration, large pore
40
what should be the gel concentration and pore size for low molecular weight samples
high gel concentration, small pore
41
does PAG have high or low resolving power?
high
42
measures the ability of an electrophoretic system to separate DNA molecules of similar sizes' effectiveness of separation
PAG
43
PAG or Agarose? can accommodate larger quantities of DNA
PAG
44
T or F Extremely turbid DNA may be recovered from PAG
F (pure)
45
T or F' PAG are stable over the wide range of pH and temperature
T
46
T or F PAG can withstand high voltage ingredients
T
47
PAG or Agarose most effective in separating DNA fragments of varying sizes from 100 bp to 25 kb
Agarose (less than 100 bp lang for PAG)
48
Agarose gels are formed by suspending dry agarose in _______
aqueous buffer
49
at what temp is agarose heated?
100 degrees celsius
50
T or F dissolved agarose gel produces a turbid mixture
F (clear, clearer than your future ganern)
51
At what temp does agarose solidify and form a gel
40-45 degrees celsius
52
T or F linear carbohydrate polymer is composed of alternating units of agarobiose, consisting of repeating units of galactose and 3,6 anhydrogalactose
F (repeating agarobiose, alternating galactose and anhydrogalactose)
53
what is the charge of DNA and RNA molecules?
negative
54
Where does DNA pieces migrate to? anode or cathode?
anode (since their charge is negative, they'll move to the positive pole)
55
A procedure that separates molecules based on their rate of movement through agarose gel under the influence of an electrical field
agarose gel elctrophoresis
56
familiarize the materials needed for agarose gel electrophoresis
electrophoresis chamber agarose gel gel casting tray buffer staining agent (dye) comb DNA ladder sample power supply
57
wire color that represents positive electrode (anode)
red
58
wire color that represents negative electrode (cathode)
black
59
material for agarose gel electrophoresis available in variety of sizes and composed of UV transparent plastic (size depends on the samples used)
gel casting trays
60
material for agarose gel electrophoresis placed in liquid agarose after it has been poured
comb
61
T or F The comb can only be placed before pouring the liquid agarose
F (before or after)
62
what does the comb produce in the hardened agarose gel?
series of wells
63
what are the wells in the agarose for?
used to load / dispense the DNA
64
Material needed for agarose gel electrophoresis solution that contains the right ions to conduct electricity
running buffer
65
What happens to DNA movement if water is used instead of running buffer
slow
66
Why is DNA movement slow if water is used instead of running buffer?
inefficient conduction of electricity because it has no ions
67
purpose of buffer
resist pH changes
68
most common buffer used in agarose gel electrophoresis
Tris buffer
69
this should be your 69th card if you're a concrete sequential learner <33
wala lang skl
70
what does TAE stand for?
Tris acetate EDTA
71
what does TBE stand for?
Tris borate EDTA
72
what does EDTA inactivate?
DNA nucleases
73
what does DNA nucleases degrade?
nucleic acids
74
buffer mostly used for the analysis of larger samples (1500 bp and higher ; generates less current)
TAE
75
buffer mostly used for smaller nucleic acid fragments
TBE
76
in what for does buffers exist in?
neutral or cationic form
77
what is the pH of TAE
8
78
what is the pH of TBE?
8
79
what is the pH of Tris Phosphate EDTA buffer?
8
80
what is the pH of Tris-citrate (TCE) buffer
7
81
what is the other name for DNA ladder
Molecular weight marker
82
consists of known DNA sizes used to determine the size of an unknown DNA sample
DNA ladder
83
T or F DNA ladder contains regularly spaced sized samples which when run on an agarose gel, it resembles a ladder
T
84
T or F Each space on a DNA ladder represents a base pair
F (Each line)
85
T or F the DNA ladder depends on the manufacturer
T
86
a set of standards used to identify the approximate size of molecule run on a gel during electrophoresis
DNA ladder
87
These are usually generated by restriction enzyme digestion of plasmid or bacteriophage DNA
DNA fragments
88
This runs alongside the DNA sample, usually on the first lane
DNA ladder
89
What should be added in the sample before being loaded on the gel?
loading dye
90
loading dye is composed of? (2)
tracking dye and density agents
91
use this to familiarize density agents
glycerol ficoll sucrose
92
What is added to the sample to make it heavier in order for it to sink in the well
loading dye (because it contains a density agent)
93
This component of loading dye provides a visual gauge of the progress of electrophoresis
tracking dye
94
Does loading dye affect the separation of the sample?
no
95
what is the other term for loading dyes? *blue text sa trans
Gel loading buffers
96
what are the two examples of tracking dye?
bromphenol blue xylene cyanol
97
A compound that can adhere to DNA and gives off fluorescent light when excited by UV light
DNA stain
98
What is the standard concentration of DNA stain used in gels
0.5 - 1 ug / ml
99
This inserts itself between the base pairs in the double helix
DNA stain
100
most commonly used DNA stain
ethidium bromide
101
safer and non-mutagenic examples of DNA stain
GelRed SYBR green
102
T or F The color of the result is dependent on the type of sample, not the stain used
F (depends on the stain)

Mol Bio Lab (12 decks)

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