Nucleic Acid (DNA Isolation) Flashcards
carries the genetic
instructions essential for the
growth, development, functioning,
and reproduction of all living
organisms
deoxyribonucleic acid
DNA is made up of
nucleotides
DNA nucleotides has three parts
2-deoxyribose
phosphate group
nitrogenous bases
DNA isolation is used for (7)
polymerase chain reaction (PCR)
DNA sequencing
gene cloning
genetic engineering
diagnosis of genetic disorders
production of vaccines
advancements in personalized medicine
why is onions used for the experiment
(1) large number of cells
(2) relatively low starch content
(3) easy to obtain
why is strawberries also used
soft and easy to pulverize
large genomes; octoploid
easy to obtain
having eight sets of chromosomes, or eight times the basic haploid chromosome number
octoploid
how many chromosomes have strawberries have in total
56 chromosomes
each cell in a strawberry contains ___ sets of chromosomes
eight
has been successfully
extracted in microgravity aboard the
International Space Station
DNA extraction
DNA extraction has been successfully
extracted in ____ aboard the
International Space Station
microgravity
DNA extraction has been successfully
extracted in microgravity aboard the
International Space Station
materials used for the experiment
50 g of Onion
Blender
Buffer solution
95% cool ethanol
buffer solution is made up of
detergent
NaCl
50 ml distilled water
equipment used for the experiment
glass rod
spatula
petri dish
hot plate
graduated cylinders
beakers
measure ___ of onion
50g
size of onion
diced (not too fine)
can also be done to cut the onion
blend
what happens to the onion if exposed to buffer solution
cell lysis
buffer solution amounts
liquid detergent
2-3 tbsp
buffer solution amounts
sodium chloride
4-5 spatula
buffer solution amounts
water
50 ml distilled water
Add the buffer solution to the onion pulp then ___
stir
Heat the mixture to ____°C for 5–10 minutes using a hot plate or water bath
40-50oC
Heat the mixture to 40–50°C for
____ minutes using a hot plate or water bath
5-10 minutes
step 1 of experiment
cell lysis using buffer solution
s the process of breaking down a cell’s membrane, which releases the cell’s contents.
cell lysis
Allow the mixture to cool to
____ temperature
room
Filter the mixture using ____ No. 1 Filter paper
Whatman No.1
Collect the ___ then set aside
filtrate
Transfer ____ mL ethanol on a Petri dish
20-30 ml
Transfer 20-30mL ____on a Petri dish
ethanol
Pour small amount of filtrate
Let mixture stand for ___ minutes
2-5 minutes
Use a _____ ____ or___ to gently spool the DNA for closer observation
glass rod
spatula
disrupts the cell membrane, releasing DNA by breaking down the lipids and proteins that maintain the membrane’s structure.
detergent
Detergent disrupts the cell membrane, releasing DNA by breaking down the ____ and ____ that maintain the membrane’s structure
lipids
process of detergent breaking down lipid and protein is similar to what
removal of grease from dishes or clothing
a biological detergent
sodium dodedcyl sulfate
SDS stands for
sodium dodecyl sulfate
further breaks down the cell membrane by disrupting the polar interactions that hold it together.
sodium dodecyl sulfate
, SDS further breaks down the cell membrane by disrupting the ____ interactions that hold it together.
polar
a key component in many laundry detergents.
sodium dodecyl sulfate
plays a crucial role in helping DNA precipitate from an ethanol solution
salt
Salt plays a crucial role in helping ____ precipitate from an ethanol solution
DNA
Salt plays a crucial role in helping DNA ____ from an ethanol solution
precipitate
Salt plays a crucial role in helping DNA precipitate from an ethanol solution by what process
neutralizing negative charges of DNA’s phosphate groups
allows the DNA strands to come closer together and clump
neutralization of - charges of DNA
carry negative charges, which are neutralized by cations like sodium.
phosphate group on DNA backbone
shield the negatively charged phosphate ends of the DNA
sodium ions
Sodium ions shield the
negatively charged phosphate ends of DNA
what happens when negatively charged phosphate ends were shielded
strands aggregate and precipitate out of the alcohol solution
Why Avoid Over-Blending or
Finely Chopping the Onion
When isolating DNA from onions, avoid blending or cutting the onion too finely, as it can slow down the filtering process.
is sufficient to break down the cell walls and membranes, efficiently releasing the DNA.
coarse blending
what will happen when you blend for too long
Blending for too long will break up the DNA fibers
is used in DNA isolation to help precipitate the DNA from the solution.
cool ethanol
ethanol is used for DNA precipitation because it enhances the process by making the
DNA less soluble than the solution
reduces the solubility of DNA even further, making the precipitation process more efficient
cooling the ethanol
also help protect the DNA from degradation during the precipitation process.
cold temperatuer
is insoluble in ethanol,
DNA
causes the DNA to clump together and separate from the rest of the solution.
cool ethanol
ethanol structure
C2H5OH
draw the structure of ethanol
check structures
temperature range helps soften the cell membrane’s phospholipids,
40-50oC temp
The 40-50°C temperature range helps soften the
cell membrane’s phospholipids
40-50oC water bath also does what
denature the proteins that could break the DNA into smaller fragments
does heating the solution below 40oC isolate the DNA?
no.
Another group heated the solution to 40˚C
for ___ minutes and successfully isolated the DNA.
17 minutes
associated onions with eternity
Egyptians
Egyptians associated onions with
eternity
were used as currency
onions
onion scientific name
Allium cepa
strawberry scientific name
Fragaria ananassa
is stored in the nucleus
DNA
DNA is stored in the ___, surrounded by the _____
___ and ___ ___ in onion cells.
nucleus
plasma membrane
cell wall
break down barriers, releasing
DNA.
detergents or lysis buffers
what process removes proteins and cellular debris
filtration
Proteins and cellular debris are removed using ____ or
mechanical filtration
proteases
Proteins and cellular debris are removed using proteases or
____ ___
mechanical filtration
are enzymes that break down proteins into smaller polypeptides or amino acids
proteases
DNA is soluble in ____but insoluble in ____,
water
alcohol
DNA is soluble in water but insoluble in alcohol, especially
with
added salts
reduces DNA solubility
cool ethanol
Cold ethanol reduces DNA solubility, causing it to
precipitate
aggregate and form visible stringy clumps due
to their long, linear, and negatively charged structure
DNA molecules
DNA molecules ___ and form visible stringy clumps due
to their long, linear, and negatively charged structure
aggregate
DNA molecules aggregate and form visible stringy clumps due
to their
long
linear
negatively charged structure
neutralize the charges on DNA, aiding in precipitation.
salts
Salts neutralize the charges on ___, aiding in precipitation.
DNA
The precipitated substance might not be pure, but
is mostly ___.
DNA
To properly evaluate the DNA yield, ____ of
the DNA is suggested
purification
example of purification process
electrophoresis
spectrophotometry
a laboratory technique used to separate DNA, RNA or protein molecules based on their size and electrical charge.
electrophoresis
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.
spectrophotometry
The whitish substance
must be ___ and ___
stringy and cohesive
challenges and solution
temperature inconsistency
heating time
Actual solution temperature
was lower than the set
50°C.
what is the solution?
increase hot plate temperature
Solution was heated for
only 10 minutes at a low
temperature.
what is the solution
Adjust
temperature and heat for an
additional 10 minutes, not
exceeding 20 minutes total.
applications of DNA
medical and health research
forensics
molecularbiology and biotechnology
agriculture
microbial studies
conservation biology
pharmaceuticals
DNA isolation application in medical and health research
genetic testing and cancer research
DNA isolation application in molecular biology and biotechnology
gene cloning and CRISPR genome editing
DNA isolation application in agricultural
crop improvement
genetically modified organisms
applications of DNA isolation in microbial studies
identification and taxonomy
metagenomics
applications of DNA isolation in in conservation biology
species idenfitication
genetic diversity analysis
DNA isolation physical interactions
chopping of onion
heat
ice bath
Chopping the onion breaks the plant’s fibrous ___which remove the cytoplasm
walls
Chopping the onion breaks the plant’s fibrous walls which remove the ____
cytoplasm
increases the rate of chemical reactions, speeding up denaturation.
heat
Heat increases the rate of chemical reactions, speeding up
denaturation
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.
ice bath
used to decrease chemical reactions
ice bath
Per heat,
____ are denatured which causes the DNA to be uncoiled.
histones
Per heat, histones are denatured which causes the DNA to be
uncoiled
chemical interactions of DNA isolation
detergent breaking down lipids
NaCl neutralizing the negative charge of phosphate backbone
precipitation of DNA using ethanol
Detergent breaks down the lipids within the ___bilayer of the cell and ___membrane, which
isolates the nuclear material (DNA) from the nucleus.
phospholipid
nuclear
isolates the nuclear material (DNA) from the nucleus.
nuclear membrane
neutralizes the negative charges on the phosphate backbone of the DNA molecule
salt
Cool ethanol causes DNA to precipitate because what does the ethanol do
pull water from DNA molecules
Cool ethanol causes DNA to precipitate because ethanol pulls water from DNA molecules, then causes
self collapse (fold in itself)
Cool ethanol causes DNA to precipitate because ethanol pulls water from DNA molecules, then causes self-
collapse (fold in itself). Thus creating
spoolable DNA
Actual DNA extraction can take up to more than ____
to ___ hours
12-30
materials used for optimal extraction of DNA
heat block
vortex mixer
microcentrifuge
