DNA ISOLATION

Relevance of DNA isolation

Isolation of DNA is often the first step before
further
analysis
DNA profiling (forensics)
Cloning
Disease diagnosis
DNA sequencing
Genetically modified organisms (GMO) agriculture,
Pharmaceutical
Environmental testing, bioterrorism

Structure of the cell

Structure of the cell
Plasma membrane and membranes of organelles
nuclear
Envelope included
DNA located in nucleus
A lot of proteins around
Mitochondrial DNA

Types of Sample

Extraction of genomic DNA

Specimen collections: right tube for

right test

Principles of DNA extraction

Cell colection
Membrane cell lysis
Protein degradation
Protein precipitation
DNA precipitation
Dissolve DNA with TE buffer or pure water

Principles
Destroy cell and nuclear
membranes, release
nuclear content (DNA
released)

Lysis buffer, SDS

Protein degradation
Protein precipitation

Proteinase K
Sodium/ammonium acetat
or extract them phenol
chloroform

DNA precipitation

Ethanol 100% in the

presence of high
salt/isopropanol

Dissolve DNA

TE pH 8 or in ultra pure
water

Storage

Phisiologic solution (TE,

H2O)

Principles DNA extraction from

Blood

DNA integrity :
Electrophoresis

Gel Electrophoresis
Apparatus

Gel Results

Gel Electrophoresis Apparatus

indwiani@yahoo.com

Gel Results

indwiani@yahoo.com

Elektroforesis Gel Agarose

Elektroforesis gel Agarose dilakukan menurut metode
yang telah dilaporkan(15). Baik gel agarose 0.8%
ataupun 1% dapat digunakan, tergantung pada ukuran
DNA yang akan dipisahkan.
Digunakan sistem larutan penyangga dengan kadar
garam rendah terbuat dari Tris-asetat 40 mM (pH 7.9)
dan sodium EDTA (larutan penyangga TAE) 2mM.
Semua proses elektroforesis diperlihatkan pada
sebuah aparatus horizontal dan dilakukan dengan
daya 100 V selama 1-3 jam.
Setelah elektroforesis dan pewarnaan dalam larutan
ethidium bromida (0.5 g/ml), gel tersebut difoto di
bawah sinar UV dengan kamera Acmel CRT M-0885D
(Polaroid), yang dilengkapi dengan filter UV jingga.