International University, Vietnam National University - HCMC 1

GENETICS LABORATORY

REPORT

PRACTICAL 4 & 5.1: EXTRACTION OF DNA FROM

PLANT CELLS & DNA QUALITY AND QUATIFICATION
Instructor : Le Minh Thong
Class: Tuesday Morning
Name: Đặng Thị Kim Sang
ID: BTBTIU17156
Date: 01/04/2019

Score: _________

1/ Explain the function of SDS buffer?

Instructor : Le Minh Thong
 International University, Vietnam National University - HCMC 2
GENETICS LABORATORY

→ SDS is an anionic detergent, which also denatures themembrane proteins and

disrupts the cell membrane. This also results in the break down of thenuclear
envelope and thereby releases the nucleoplasm. SDS also helps in inhibition of
nucleases.
2/ Function of Amonium acetate?
→The salt, in this case ammonium acetate, helps separate the proteins from the
DNA. It helps us get close to 100% DNA and not the histones (or proteins) that
DNA is wrapped around.
3/ What is the color of DNA after extration?
In outside, DNA after extraction almost are colorless. The yellow region in the gel
appears by polyphenol.Depending on the
species, plant cells can also contain lots of
contaminats that have the potential to
inhibit downstream reactions.
Polysaccharides, polyphenols, and tannins
are all common in the plant samples and
are usually difficult to remove if allowed
to persist all the way through the
extraction process.

We have the negative end and the positive end. We're going to load our DNA at
the negative end because DNA samples are negatively charged .So those represent
the wells and the electrophoresis reaction is occurring, the negative current here is
going to push the samples toward the positive end.And you may end up with
Instructor : Le Minh Thong
 International University, Vietnam National University - HCMC 3
GENETICS LABORATORY

something that looks like the image. Each of

these lines represents a band of DNA. All the
pieces of DNA in this band are the same size,
and you can see in the picture everything this
distance is the same size. Even though we
have two different samples are approximately
the sample size fragments. Those fragments
are the smallest are going to move the farthest
and those fragments that are the largest are
not going to travel as far. There's reason for
that and that deals with how agarose has
made. Depending on the concentration you
have of agarose . It's kind of like a mesh I like
to use the analogy that it's like the trees in the
forest are really close together or the trees in
the forest are farther apart. If you have a
really far apart forest the bigger pieces will be able to move a little bit farther.So if
you are on a horse in the forest that horse is going to be able to move fairly easily
through the forest. If you are in this forest where the trees are much closer
together on your on that same horse, you are going to move a lot more slowly
.And so if you have a dog with you while you are riding through the forest on a
horse , in the forest farther apart the dog is going to move even more quickly than
the horse because it's smaller and down, it is going to move much more quickly in
the forest where the trees are much closer. And so we have our large fragment in
closer region and our small fragment in farther region, then the gel in this is less
concentrated that horse is going to move a little more father down the band or
down the gel and the dog is the smaller fragments are going to move very far .
And so that's the difference you can see between your two different
concentrations of gel and in general how this works. In the police world , they
would be able to compare this if it was a paternity test and try to find matching
bands .

4/How do you know you extraction contain DNA?

Instructor : Le Minh Thong

 International University, Vietnam National University - HCMC 4
GENETICS LABORATORY

In order to extract DNA from leaf, we must use

a liquid “extractionbuffer”. The extraction
buffer is easy to make and contains
dishwashingliquid (soap), salt and water. Each
component of the extraction bufferperforms a
specific function. The soapbreaks up the
phospholipid bilayersof the cell membrane,
organelle membranes and nuclear membrane.
The saltbreaks up protein chains that may be
bound to the DNA, such as plant pigments and
tannins (the proteins that make coffee and tea
taste bitter). In addition to the extraction buffer,
we will be using 100% ethanol or isopropanol,
both of which are types of alcohol. Once we
mix the smashed leaf with the extraction buffer, we will use this alcohol to
precipitate the DNA. Precipitation of DNA occurs because DNA is not soluble in
ethanol or isopropanol. The colder the alcohol, the less soluble the DNA will be in
it. Therefore, we know extraction contain DNA when the last step appear
precipitate.

Instructor : Le Minh Thong