DNA ISOLATION

PURPOSE :
To know simple method oII Iruits DNA isolation
To know soap or detergent eIectiIity used Ior DNA isolation

BASIC THEORY
DNA is a nuclead acid which contain some genetics materials and have
Iunction to arrange physical grow every liIe Iorm in cellular Iorm. DNA can be Iound
in the nucleus, mitocondria, and chlorolpast. The diIIerence between each other is in
nucleus lineary Iorm and close asociated with histon protein, meanwhile
mitochondria and chloroplast DNA are sirculary Iormed and doesn`t asociated with
histon protein. Beside it there are also other diIIerence between it, mitochondria and
chloroplast DNA only inherit traits which only come Irom the mother side. In the
nucleus DNA heredity patern inherit both oI it parental, which means Iather side and
mother side. II we see it Irom the organism, the diIIerence between procariotic DNA
and eucariotic DNA are same with the diIIerence between nucleus and mitochondria,
that is in eucariotic lineary Iormed and have histon proteins meanwhile in procariotic
are cilculary Iormed doesn`t have histon proteins.
According the structure, DNA consist oI some components there are, sugar on
the pentose group (deoxyribose), phosIat group, and base nitrogenes. The base
nitrogenes which composed it devided into 2 type there are purin and pirimidin. Purin
base are devided into 2 type there are adenin (A) and guanin (G) which both oI it
have double ring structure. Pirimidin base also devided into 2 type there are cytocin
(C) and timin (T) which have single ring structure. When guanin bounded with
cytocin, it will Iorming 3 hydrogen bound meanwhile, when admin bounded with
timin it wiil only Iorming 2 hydrogen bound. A single building blick oI DNA consist
oI a sugar, phosIat group, and a pair oI base nytrogene. It will be Iorming double
spiral (double helix) and called nucleotide.
When we learn or modoIication oI DNA, we need a method in order to make
it easy and 1 oI methods are with doing DNA isolation in the chromosome. DNA
isolation is a method to separating components within DNA, it using sentryIugal
theory. Basically we can used centryIugal machine to observe it, iI we use centryIugal
machine there are steps to do it:
O Tissue isolation
O Lysising membrane and cell wals
O Extracting to the solution
O PuriIication
O Precipitation

The principles in conducting DNA isolation there are two namely
centriIugation and precpitaion. The main principle is to
separate substances by centriIugation speciIic gravity oI the molecule by providing
a centriIugal Iorce so that the heavier substance will, while the lighter substance will
be located at the top. CentriIugation technique was perIormed in a machine with
variable speed, Ior example 2500 rpm(rotation per minute) or 3000 rpm
Based Irom the steps i`ve mention it above, the steps to do DNA isolation Iirst
is tissue isolation. Tissue isolation is a Iirst step to do it, it choose the material step.
second is lysising membraneis lysising membrane and cell wall, the main Iunction oI
this step is to break the cell membrane and cell wall in order to make the DNA out.
The third step is extracting the solution, extracting solution main purpose is to get the
main extract oI the solube. Fourth step is puriIication the puriIication have main
objection to clean the solube Irom the unwanted thing. FiIth step is precipitation,
precipitation is last step oI it, it devide the solube by the diIIerence oI the sulube
viscosity and make the solube homogen.
Isolation oI DNA by centriIugation technique would precipitate the DNA. In
order Ior theisolation oI pure DNA that is not mixed with other molecules in the
process oI isolationthen mixed a variety oI solutions. A solution which serves as
resuspention Iusion backpellets that have been Iormed with mixed solution.
Additionally solution A also serves as a buIIer and chelating agent. The selection
buIIer is viewed Irom the ability to buIIergenerate electric current. Solution B
consisting oI SDS and NaOH solution serves as lysising
NaOH alone can mendenaturasi proteins. While the solution oI C have
Iunction to renaturate back. 70 ethanol used in the isolation process serves to
remove salt deposits because Na is positively charged and negatively charged
DNA. DdH
2
O solution is added to Iunction so that the resulting DNA precipitate
obtained with high enough concentrations. The vortex solution to Iacilitate the
bacteria suspension. The DNA that have been precipitated by the addition oI the kana
lysis buIIer soution oI the kana lysis buIIer solution.








.
PRACTICE METHODS
Tools and Equipment
O Beaker glass
O niIe
O Mixer
O Filter (tissue or cotton)
O Blender
O Spatula
O React tube with the shelI
O Tomato, papaya, and pear Iruit
O Detergent (surI, rinso)
O Pinch soap
O Salt
O Cold ethanol 96 (ethanol with ice)
O Aquadest
Schematic work
Disolve the detergent and pinch soap

Blend Iruit with aquadest

Mix each mixture

Add it with salt

Filter it twice

Add the Iilter result with cold ethanol

Observe the process DNA occurity















OBSERVATION RESULT
et Mixture BeIore droped with
ethanol
AIter droped DNA appear
1 Tomato rinso
2,3 ml ethanol
1,92 ml
Single layer yellow
muddy colored
3 layers (Irom
bottom)
Tomato
DNA
Rinso and
ethanol
113 seconds
2 Pear rinso 2,1
ml ethanol 1,75
One layer, transparent
and soIt yellow colour
4 layers (Irom
bottom)
Pear
Ethanol
DNA
detergent
46 seconds
3 Papayarinso 2,6
mlethanol 2,16
ml
Clearly orange consist
oI 1 layer
3 layers
Rinso and
ethanol
DNA
Papaya esence
another part
oI it

76 seconds
4 Tomato surI 2,3
ml ethanol 2,9
ml
The color is yellow
clear only a layer
3 layers
SurIethanol
DNA
Pear essence
54 seconds
5 Pear surI 2,1 ml
ethanol 2,5 ml
The colour is yellow
clear and consist a
3 layers
SurIethanol
68 seconds
layer DNA
Pear essence
6 PapayasurI 2,2
mlethanol 1,82
ml
The colour is orange
pale and consist a
layer
3 layers
SurIethanol
DNA
Papaya
essence
85 seconds
















EXPLANATION
DNA isolation can do it by various way one oI the is by using adding some
liquid to the Iruit essence, the Iruiit essence can be goten by pulverize the Iruit by
using blender about 40 seconds until 1 minute. In the that interval is the best time to
pulverize the Iruits to prevent the Iruit DNA to break. The main purpose pulverize
itselI is to break the cell membrane, cell wall, and core membrane in order to make th
DNA can escape Irom it. To brek the membrane also can using the detergent .
The main Iunction oI the various liquid which added to the the object is Ior
buIIer oI the Iruit essence, the buIIer have main Iunction to make clear the liquid. The
buIIer chosen Ior the practice is the liquid which have ability to generate electirIy, so
in this case the buIIer chosen are ethanol and salt (NaCl) which both oI it have the
ability to generate the electriIy
Based on the observation result we did 3
rd
experiment by using papayarinso
2,6 ml ethanol 2,16 ml and added with salt (NaCl) we get result it change Irom
Clearly orange consist oI 1 layer transIorm into 3 layers oI Iruit essence which iI we
count it Irom bottom are rinso and ethanol, DNA, and papaya essence and that apear
in the 76 seconds. It can be happened because the salt (NaCl) consinst oI ion Na

and
when the salt bonded with phospat give the negative bound pole and the DNA will
gather. Meanwhile the Iunction oI the Ethanol is to do DNA precipitate the liquid
which DNA has gathered able to selI separated Irom the liquid and make layers whic
able to identiIied the materials.
Meanwhile the other group are use the diIIerent materials get the result oI the
observation whih i`ll mention it below :
O Group 1 which using Tomato rinso 2,3 ml ethanol 1,92 ml Ior they
experiment get the result, beIore droped with ethanol it have yellow colour
and single layer. AIter it dropped with the ethanol it change into 3 layers
which iI it counted Irom bottom consist oI tomato essence, DNA, rinso and
ethanol in 113 seconds
O Group 2 which using Pear rinso 2,1 ml ethanol 1,75 ml Ior they
experiment get the result, beIore it droped with ethanol it have single layer,
transparent and soIt yellow colour. AIter it dropped with ethanol it change
into 4 layers which iI it counted Irom the bottom consist oI pear, DNA,
ethanol, and rinso. And it appear in 46 sseconds
O Group 4 which using Tomato surI 2,3 ml ethanol 2,9 ml Ior they
experiment get the result, beIore it dropped with ethanol it have clear yellow
colour and consist single layer. AIter it dropped with ethanol it transIorm into
3 layers which iI it counted Irom the bottom consist oI tomato , DNA, etanol
and surI. And it appear in 54 seconds
O Group 5 which using Pear surI 2,1 ml ethanol 2,5 ml Ior they experiment
get the result, beIore it dropped with ethanol it have clear yellow and have
single layer. AIter it dropped with ethanol it transIorm into 3 layers which iI it
counted Irom the bottom it consist pear essence, DNA, surI and ethanol. And
it appear in 68 seconds
O Group 6 which using PapayasurI 2,2 mlethanol 1,82 ml Ior they
experiment get the result, beIore it dropped with ethanol it pale orange
coloured and have single layer. AIter it dropped with ethanol it transIorm into
3 layers which iI it counted Irom bottom it consist papaya essence, DNA, surI
nd ethanol/ and it appear in 85 seconds





CONCLUTION
Based Irom the observation which we did we can get some conclution there are:
O The diIIerence between each Iruits to show the eIIect oI the mixture liquid are
various, it depend on the Iruit and the detergent. The Iruit determine the
duration oI the DNA will appear because the water which consist within it, the
more water within the Iruits the more time it needed.
O The Iunction oI the various mixture oI liquid which added to the Iruit essence
are: ethanol have Iunction to hasten the process oI DNA occurity and sent out
the salt reside, salt (NaCl) have Iunction to devide the electron within the Iruit
essence in order to keep the Ph. The detergent Iunction is to break the cell
membrane, cell wall, and core cell in order to make DNA can expeled.
O In my opinion the Iruit which contain most oI water within it, is tomato
because Irom the all result oI the group observation it showing iI it need
longest time to DNA appear.










REFERENCES
Campbell,Reece,Mitchell. 2004. BIOLOGI JILID I Edisi kelima. Jakarta : Penerbit
Erlangga
Genetic Team. 2011. Genetic Labwork Module. Jember: Jember University.
Istanti, Annie. 1999. Biologi Sel. Malang: jurusan Biologi FMIPA UM.
Rizqiawan, angga, 2011. Isolasi DNA ~ SAINS WORLD.
http://duniasains-rizqy.blogspot.com/2011/11/isolasi-dna.html accesed in 15-11-
2011
Sesyuhada. 2009. ISOLASI DNA Seisyuhada`s Weblog
http://seisyuhada.wordpress.com/2009/11/20/isolasi-dna/ accesed in 15-11-
2011