COMPLETE REPORT GENETIC
DNA ISOLATION
Which made by:
Ilham
Nur
1414440012
ICP B Biology
BIOLOGY DEPARTMENT
MATHEMATIC AND SCIENCE FACULTY
STATE UNIVERSITY OF MAKASSAR
2016
RATIFICATION
PAGE
Complete report of genetic practicum with the
title “DNA ISOLATION”
that arranged by:
Name : Ilham Nur
ID :
1414440012
Class : ICP B
Group :
3
has been checked
by assistant and assistant coordinator, so this report was accepted
Makassar
05 January 2017
Asistant Coordinator Asistan
Ferry Irawan ,S.Pd Ferry
Irawan ,S.Pd
CHAPTER I
INTRODUCTION
A.
Background
Deoxyribonucleic
acid or DNA is a chemical compound that is most important in life. DNA
creature is a compound that contains the genetic information of living beings
from one generation to the next. The entire DNA in a cell will form the genome.
Genome includes genes that are part of the functional and non-functional in the
cells of organisms. DNA genome includes genes danintergen. DNA is a nucleic acid that contains
the genetic material and serves to regulate the biological development of all
forms of life in a cell.
DNA
found in the nucleus, mitikondria, and chloroplasts. Third difference is linear
and shaped nucleus DNA is closely associated with histone proteins, while the
DNA of mitochondria and chloroplasts circular shaped and are not associated
with histone proteins. In addition mitochondrial DNA and chloroplast has a
characteristic, which is only inherited traits come from the mother. While
nuclear DNA has a pattern of inheritance from both parents. Judging from the
organism, the structure of prokaryotic DNA and histone proteins do not have a
circular shape, while the form of linear eukaryotic DNA and have a histone
protein.
DNA isolation is a step in studying DNA.
One
prinsipisolasi DNA is by centrifugation. Centrifugation a mixture
untukmemisahkan techniques based on molecular weight components. Molecules that
have a large molecular weight will be on the bottom of the tube and the lighter
molecules will be at the top of the tube. Centrifugation results will show two
kinds of separate fractions, namely the supernatant at the top and pellet on
the bottom
B .
Purpose
The purpose on this practicum that to know how to
isolated DNA by using the simple
material.
C.
Benefit
The student can more know how to isolate DNA by the
simple practicum at Isolate DNA
CHAPTER II
PREVIEW OF
LITERATURE
In eukaryotic cells
including plants and animals are the biggest part of the DNA in the nucleus is
an organelle that is separated from the cytoplasm by a membrane. The nucleus
consists of 90% of total cellular DNA. Residual DNA are organelles such as
mitochondria and chloroplasts. Because the DNA found in the nucleus, it is necessary
for cell pelisisan method to harvesting the cells. Wherein the method is part
of the DNA isolation method Separation
of DNA from other cellular material is significant and requires penyallinan DNA
into RNA and translation of RNA into proteins takes place in compartment
(space) are different, namely in a row in the nucleus and cytoplasm (Beckingham , 2005)
There is a double
membranous organelles in the cytoplasm, including the mitochondria in both
plants and animals. Therefore it is necessary to isolate DNA in plants and
animals to learn and study the DNA of plants and animals. Eukaryotic cells have
more DNA, complete with other components. DNA of plants and animals stored in
nucleusyang encased by a membrane. Isolation of DNA is the right step for
studying DNA. In principle there are two, namely the centrifugation and
presipitasi.Sentrifugasi is a technique for separating mixtures based on
molecular weight components. Molecules that have a large molecular weight will
be on the bottom of the tube and the lighter molecules will be at the top of
the tube. Centrifugation results will show two kinds of separate fractions,
namely the supernatant at the top and pellets at the bottom. Precipitation is a
step that is carried out to precipitate a component of the (Chhabra, 2013)
DNA in living organisms
can be isolated simply. DNA isolation can simply be done by breaking the cell
wall, the plasma membrane and nuclear membrane either mechanically or
chemically. Isolation of DNA is a technique used to obtain pure DNA, ie without
the protein and RNA from a cell in the network. Mechanically breaking the cell
wall can be done with pemblenderan or grinding using a mortar and pistil. While
chemically can be done with the detergent. The addition of liquid soap and salt
is to lyse the nuclear membrane to release the contents of the cell nucleus
which contains DNA (Parvathi, 2009)
DNA Isolation can be
done through the stages include: preparation of cell extracts, purification of
DNA from ekstrsk cells and DNA precipitation. Although DNA isolation can be
done in various ways, but in any kind or plant part may give different results,
this is due to compounds called polyphenols and polysaccharides in high
concentrations can inhibit DNA purification. If the sample DNA isolation is
done with fruit, the water content in each of the different pieces, can give
different results as well. The higher the water content, the cells are
dissolved in the extract will be less, so that the DNA terpretisipasi also be (Parvathi, 2009).
Each breed or events
described in advance is determined by genes located on autosomes. In addition
to known genes as well as genes that are called genes strung sex. The chain of
events called the genitals or in the English called Sex. Usually the dominant gene
showed effects on individual men and women, or male or female. New in the
homozygous recessive state dominant influence it will not appear in the
phenotype. If we put the right hand or the left right on a pedestal that there
is a horizontal line such that the tip of the ring finger touching the line,
then we can know whether our index finger will be longer or is shorter than the
ring finger. In most people the index finger will not reach the line that
identifies that some individuals may have the same absolute gene content of
inactivity chromosome creates a sub-population to distinguish the content of
active genes (Parvathi, 2009)
The addition of
detergent in the isolation of DNA can cause damage to the cell membrane,
through the bonds formed through the hydrophobic detergent with proteins and
fats in the membrane forming compounds "protein-detergent lipid
complex". The compounds can be formed as proteins and lipids have a
hydrophilic and hydrophobic end, as well as detergents, so as to form a chemical
bond (Ramesh,
2014)
CHAPTER III
EXPERIMENT METHOD
A.
Time and Place
Day
/ Date : Friday, January 6th
2016
Time : 10.00 am – 11.30 am
Place :
Biology Laboratory 2nd Floor of the east, Faculty of Mathematic and Sience, state University of
Makassar.
B. Tools
and Materials
1. Tools
a.
Reaction
tube
b.
Knife
c.
Pipette
d.
Beaker
class
e.
Mortal
2. Materials
a.
Pear
b.
Drogon
Fruit
c.
Kiwi
d.
Strawberry
e.
Detergent
f.
Salt
g.
Ethanol
C.
Work
Procedure
fruit
crush using mortar then added aquades and detergent until homogen
|
Choose
the good for the friut and cut by uing knife,
|
Filter
solution of Friut and detergent into beaker glass then pull into reaction
tube
|
Added
NaCL and Cold Ethanol and turn on the Stopwatch wait untill reaction
|
CHAPTER IV
OBSERVATION RESULT AND DISCUSSION
A.
Observation Result
a.
Kiwi
b.
Dragon Fruit
c.
Pear
d. Strawberry
B.
Discussion
The
main principle in the isolation of DNA, there are three namely the destruction
(lysis), DNA extraction or separation of solid materials such as cellulose and
proteins, as well as DNA
purification. In addition there are two principles in conducting DNA isolation,
ie centrifugation and precipitation. The main principle is centrifuged to
separate the substance by weight types of molecules by providing a centrifugal
force so that the heavier substances will be at the bottom, while the lighter
substances will be located above. Centrifugation techniques are performed in a
machine called a centrifuge machines at varying speeds.
In
this experiment, we performed the isolation of DNA derived from two types of
fruit which has a different water content. The purpose of this experiment is to isolate or separate the DNA from
plants. The method This
experiment was conducted in the destruction (lysis) and extraction. The first
treatment given to the fruit is peeled and cut menajadi smaller size, it is
done so at the time of the destruction of the fruit is crushed into particles -
smaller particles. The next stage is to add a mixture of detergent with water,
it aims to damage the cell membrane of the fruit. The destruction of the cell
membrane occurs due to the chemical bond formed between detergent with
substances that exist on the fruit. After the addition of detergent next stage
of penambahana salt as much as 1 spatula to each tube containing a mixture of
fruit and detergent, the purpose of the addition of salt is to facilitate the
separation of the threads of DNA from the mixture so that the threads will be
easily observed. This happens because the Na + in salt can form a bond to the
negative pole of DNA.Setelah phosphate bonding solution is added salt solution
were homogenized on a vortex machine. From the experimental results
obtained DNA isolation that is, differences in color on each tube containing a
mixture of fruits, detergents, salts, and ethanol.Perubahan colors that occur
due to differences in the composition of the DNA contained in each fruit.
Besides the striking difference from the sixth solution is there are a lot or a
little sediment at the nucleic acid.
This
method does not require the cost of more expensive compared to using the kit.
In addition, the advantages of this is the extraction of DNA bands yangdiproleh
thicker when dibandinglan with phenol extraction method and without phenol.
However, the results with this method there are ribbon smear and DNA generated
fewer than extraction using a kit. Constraints common in the extraction of CTAB
is the inhibitor on the host, the low concentration of vius and influence how
and long storage time.
CHAPTER V
CONSLUSSION ANG SUGGESTION
a. Conclussion
DNA isolation from the
experiments that we can know the method or how to isolate DNA from a fruit used
as a sample. Namely through how the destruction (lysis), extraction, and
purification. The use of different detergents in the experiments conducted to
determine the detergent in the form of what are the most active in destroying
the cell membrane in each sample buah.Setelah conducted the experiment turned
out to produce DNA coarse powder detergent, more visible and more white
sediment.
b. Suggestion
At all was good. Nothing
suggest anymore.
BIBLIOGRAPHY
Beckingham kathleen m. 2005. Drosophila melanogaster
- the model organism of choice for the complex biology of multi-cellular
organisms. Gravitational and space biology journal. Vol 18(2). Department of
biochemistry and cell biology, ms-140, rice university, houston. Texas
Chhabra,
Ria. et all. 2013. Organically Grown Food Provides Health Benefits to
Drosophila melanogaster. International Journal of Advanced Research. Volume 8,
Issue 1 Clark High School, Plano, Texas, United States of America, 2 Department
of Biological Sciences, Southern Methodist University. Dallas, Texas, United
States of America
Parvathi, deepa v, et all. 2009. Wonder animal model
for genetic studies - drosophila melanogaster its life cycle and breeding
methods. Journal of medicine. Vol. Ii, issue 2. Department of human genetics
sri ramachandra university porur. Chennai
Ramesh,
B.Y, et all, 2014. Carbohydrate and protein are an attribute to enhance the
life-history determinants in Drosophila. International Journal of Advanced
Research. ISSN 2320-5407. Volume 2. Issue 1. Centre for Applied Genetics,
Bangalore University, Bangalore-560 056, Karnataka, India
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