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Biomolecules > Nucleic Acid (DNA Isolation) > Flashcards

Nucleic Acid (DNA Isolation) Flashcards

1
Q

carries the genetic
instructions essential for the
growth, development, functioning,
and reproduction of all living
organisms

A

deoxyribonucleic acid

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2
Q

DNA is made up of

A

nucleotides

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3
Q

DNA nucleotides has three parts

A

2-deoxyribose
phosphate group
nitrogenous bases

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4
Q

DNA isolation is used for (7)

A

polymerase chain reaction (PCR)
DNA sequencing
gene cloning
genetic engineering
diagnosis of genetic disorders
production of vaccines
advancements in personalized medicine

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5
Q

why is onions used for the experiment

A

(1) large number of cells
(2) relatively low starch content
(3) easy to obtain

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6
Q

why is strawberries also used

A

soft and easy to pulverize
large genomes; octoploid
easy to obtain

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7
Q

having eight sets of chromosomes, or eight times the basic haploid chromosome number

A

octoploid

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8
Q

how many chromosomes have strawberries have in total

A

56 chromosomes

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9
Q

each cell in a strawberry contains ___ sets of chromosomes

A

eight

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10
Q

has been successfully
extracted in microgravity aboard the
International Space Station

A

DNA extraction

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11
Q

DNA extraction has been successfully
extracted in ____ aboard the
International Space Station

A

microgravity

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12
Q

DNA extraction has been successfully
extracted in microgravity aboard the

A

International Space Station

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13
Q

materials used for the experiment

A

50 g of Onion
Blender
Buffer solution
95% cool ethanol

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14
Q

buffer solution is made up of

A

detergent
NaCl
50 ml distilled water

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15
Q

equipment used for the experiment

A

glass rod
spatula
petri dish
hot plate
graduated cylinders
beakers

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16
Q

measure ___ of onion

A

50g

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17
Q

size of onion

A

diced (not too fine)

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18
Q

can also be done to cut the onion

A

blend

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19
Q

what happens to the onion if exposed to buffer solution

A

cell lysis

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20
Q

buffer solution amounts

liquid detergent

A

2-3 tbsp

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21
Q

buffer solution amounts

sodium chloride

A

4-5 spatula

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22
Q

buffer solution amounts

water

A

50 ml distilled water

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23
Q

Add the buffer solution to the onion pulp then ___

A

stir

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24
Q

Heat the mixture to ____°C for 5–10 minutes using a hot plate or water bath

A

40-50oC

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25
Q

Heat the mixture to 40–50°C for
____ minutes using a hot plate or water bath

A

5-10 minutes

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26
Q

step 1 of experiment

A

cell lysis using buffer solution

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27
Q

s the process of breaking down a cell’s membrane, which releases the cell’s contents.

A

cell lysis

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28
Q

Allow the mixture to cool to
____ temperature

A

room

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29
Q

Filter the mixture using ____ No. 1 Filter paper

A

Whatman No.1

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30
Q

Collect the ___ then set aside

A

filtrate

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31
Q

Transfer ____ mL ethanol on a Petri dish

A

20-30 ml

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32
Q

Transfer 20-30mL ____on a Petri dish

A

ethanol

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33
Q

Pour small amount of filtrate
Let mixture stand for ___ minutes

A

2-5 minutes

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34
Q

Use a _____ ____ or___ to gently spool the DNA for closer observation

A

glass rod
spatula

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35
Q

disrupts the cell membrane, releasing DNA by breaking down the lipids and proteins that maintain the membrane’s structure.

A

detergent

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36
Q

Detergent disrupts the cell membrane, releasing DNA by breaking down the ____ and ____ that maintain the membrane’s structure

A

lipids

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37
Q

process of detergent breaking down lipid and protein is similar to what

A

removal of grease from dishes or clothing

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38
Q

a biological detergent

A

sodium dodedcyl sulfate

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39
Q

SDS stands for

A

sodium dodecyl sulfate

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40
Q

further breaks down the cell membrane by disrupting the polar interactions that hold it together.

A

sodium dodecyl sulfate

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41
Q

, SDS further breaks down the cell membrane by disrupting the ____ interactions that hold it together.

A

polar

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42
Q

a key component in many laundry detergents.

A

sodium dodecyl sulfate

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43
Q

plays a crucial role in helping DNA precipitate from an ethanol solution

A

salt

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44
Q

Salt plays a crucial role in helping ____ precipitate from an ethanol solution

A

DNA

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45
Q

Salt plays a crucial role in helping DNA ____ from an ethanol solution

A

precipitate

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46
Q

Salt plays a crucial role in helping DNA precipitate from an ethanol solution by what process

A

neutralizing negative charges of DNA’s phosphate groups

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47
Q

allows the DNA strands to come closer together and clump

A

neutralization of - charges of DNA

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48
Q

carry negative charges, which are neutralized by cations like sodium.

A

phosphate group on DNA backbone

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49
Q

shield the negatively charged phosphate ends of the DNA

A

sodium ions

50
Q

Sodium ions shield the

A

negatively charged phosphate ends of DNA

51
Q

what happens when negatively charged phosphate ends were shielded

A

strands aggregate and precipitate out of the alcohol solution

52
Q

Why Avoid Over-Blending or
Finely Chopping the Onion

A

When isolating DNA from onions, avoid blending or cutting the onion too finely, as it can slow down the filtering process.

53
Q

is sufficient to break down the cell walls and membranes, efficiently releasing the DNA.

A

coarse blending

54
Q

what will happen when you blend for too long

A

Blending for too long will break up the DNA fibers

55
Q

is used in DNA isolation to help precipitate the DNA from the solution.

A

cool ethanol

56
Q
A
57
Q

ethanol is used for DNA precipitation because it enhances the process by making the

A

DNA less soluble than the solution

58
Q

reduces the solubility of DNA even further, making the precipitation process more efficient

A

cooling the ethanol

59
Q

also help protect the DNA from degradation during the precipitation process.

A

cold temperatuer

60
Q

is insoluble in ethanol,

A

DNA

61
Q

causes the DNA to clump together and separate from the rest of the solution.

A

cool ethanol

62
Q

ethanol structure

A

C2H5OH

63
Q

draw the structure of ethanol

A

check structures

64
Q

temperature range helps soften the cell membrane’s phospholipids,

A

40-50oC temp

65
Q

The 40-50°C temperature range helps soften the

A

cell membrane’s phospholipids

66
Q

40-50oC water bath also does what

A

denature the proteins that could break the DNA into smaller fragments

67
Q

does heating the solution below 40oC isolate the DNA?

A

no.

68
Q

Another group heated the solution to 40˚C
for ___ minutes and successfully isolated the DNA.

A

17 minutes

69
Q

associated onions with eternity

A

Egyptians

70
Q

Egyptians associated onions with

A

eternity

71
Q

were used as currency

A

onions

72
Q

onion scientific name

A

Allium cepa

73
Q

strawberry scientific name

A

Fragaria ananassa

74
Q

is stored in the nucleus

A

DNA

75
Q

DNA is stored in the ___, surrounded by the _____
___ and ___ ___ in onion cells.

A

nucleus
plasma membrane
cell wall

76
Q

break down barriers, releasing
DNA.

A

detergents or lysis buffers

77
Q

what process removes proteins and cellular debris

A

filtration

78
Q

Proteins and cellular debris are removed using ____ or
mechanical filtration

A

proteases

79
Q

Proteins and cellular debris are removed using proteases or
____ ___

A

mechanical filtration

80
Q

are enzymes that break down proteins into smaller polypeptides or amino acids

A

proteases

81
Q

DNA is soluble in ____but insoluble in ____,

A

water
alcohol

82
Q

DNA is soluble in water but insoluble in alcohol, especially
with

A

added salts

83
Q

reduces DNA solubility

A

cool ethanol

84
Q

Cold ethanol reduces DNA solubility, causing it to

A

precipitate

85
Q

aggregate and form visible stringy clumps due
to their long, linear, and negatively charged structure

A

DNA molecules

86
Q

DNA molecules ___ and form visible stringy clumps due
to their long, linear, and negatively charged structure

A

aggregate

87
Q

DNA molecules aggregate and form visible stringy clumps due
to their

A

long
linear
negatively charged structure

88
Q

neutralize the charges on DNA, aiding in precipitation.

A

salts

89
Q

Salts neutralize the charges on ___, aiding in precipitation.

A

DNA

90
Q

The precipitated substance might not be pure, but
is mostly ___.

A

DNA

91
Q

To properly evaluate the DNA yield, ____ of
the DNA is suggested

A

purification

92
Q

example of purification process

A

electrophoresis
spectrophotometry

93
Q

a laboratory technique used to separate DNA, RNA or protein molecules based on their size and electrical charge.

A

electrophoresis

94
Q

is a method to measure how much a chemical substance absorbs light by measuring the intensity of light as a beam of light passes through sample solution.

A

spectrophotometry

95
Q

The whitish substance
must be ___ and ___

A

stringy and cohesive

96
Q

challenges and solution

A

temperature inconsistency
heating time

97
Q

Actual solution temperature
was lower than the set
50°C.

what is the solution?

A

increase hot plate temperature

98
Q

Solution was heated for
only 10 minutes at a low
temperature.

what is the solution

A

Adjust
temperature and heat for an
additional 10 minutes, not
exceeding 20 minutes total.

99
Q

applications of DNA

A

medical and health research
forensics
molecularbiology and biotechnology
agriculture
microbial studies
conservation biology
pharmaceuticals

100
Q

DNA isolation application in medical and health research

A

genetic testing and cancer research

101
Q

DNA isolation application in molecular biology and biotechnology

A

gene cloning and CRISPR genome editing

102
Q

DNA isolation application in agricultural

A

crop improvement
genetically modified organisms

103
Q

applications of DNA isolation in microbial studies

A

identification and taxonomy
metagenomics

104
Q

applications of DNA isolation in in conservation biology

A

species idenfitication
genetic diversity analysis

105
Q

DNA isolation physical interactions

A

chopping of onion
heat
ice bath

106
Q

Chopping the onion breaks the plant’s fibrous ___which remove the cytoplasm

A

walls

107
Q

Chopping the onion breaks the plant’s fibrous walls which remove the ____

A

cytoplasm

108
Q

increases the rate of chemical reactions, speeding up denaturation.

A

heat

109
Q

Heat increases the rate of chemical reactions, speeding up

A

denaturation

110
Q

In the experiment, it is instructed to wait until the sample is at room temperature. However, an ___ ____ can also
be used to decrease chemical reactions, a countermeasure to the heat applied.

A

ice bath

111
Q

used to decrease chemical reactions

A

ice bath

112
Q

Per heat,
____ are denatured which causes the DNA to be uncoiled.

A

histones

113
Q

Per heat, histones are denatured which causes the DNA to be

A

uncoiled

114
Q

chemical interactions of DNA isolation

A

detergent breaking down lipids
NaCl neutralizing the negative charge of phosphate backbone
precipitation of DNA using ethanol

115
Q

Detergent breaks down the lipids within the ___bilayer of the cell and ___membrane, which
isolates the nuclear material (DNA) from the nucleus.

A

phospholipid
nuclear

116
Q

isolates the nuclear material (DNA) from the nucleus.

A

nuclear membrane

117
Q

neutralizes the negative charges on the phosphate backbone of the DNA molecule

A

salt

118
Q

Cool ethanol causes DNA to precipitate because what does the ethanol do

A

pull water from DNA molecules

119
Q

Cool ethanol causes DNA to precipitate because ethanol pulls water from DNA molecules, then causes

A

self collapse (fold in itself)

120
Q

Cool ethanol causes DNA to precipitate because ethanol pulls water from DNA molecules, then causes self-
collapse (fold in itself). Thus creating

A

spoolable DNA

121
Q

Actual DNA extraction can take up to more than ____
to ___ hours

A

12-30

122
Q

materials used for optimal extraction of DNA

A

heat block
vortex mixer
microcentrifuge

Biomolecules (23 decks)

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