Beruflich Dokumente
Kultur Dokumente
Electrophoresis of DNA
Natalie B. Nisce
Department of Food Science and Nutrition, College of Home Economics, University of the Philippines Diliman,
Quezon City, Philippines
ABSTRACT The following experiment describes the process and principles behind extracting DNA from live shrimps
and assessing the DNA extracts concentration and purity, based on the detection of its basic unit, nucleic acid. This
experiment aims to obtain an extract with high nucleic acid purity and concentration. The extraction process involves
cell lysis, the denaturing of proteins and destruction of protein-DNA complexes. The purity of the nucleic acid obtained
was assessed using a double beam UV-vis spectrophotometer at wavelengths 260nm and 280nm. The recorded
absorbance values were 0.6598 and 0.4757 for 260nm and 280nm, respectively. The % nucleic acid of the extract was
10% and the concentration of DNA was computed to be 3.299
. To further evaluate the purity of the extracted
DNA, the obtained sample was subjected to agarose gel electrophoresis. However, due to possible old reagents, the
gel did not impart clear bands. Apart from the agarose gel electrophresis, it can be concluded that the methods used in
this experiment are significant in the extraction and overall study of DNA.
INTRODUCTION
All living organisms contain the genetic carriers,
DNA. With the exception of identical siblings, each
organism carries a different set of DNA, making each
one unique from the other. This is because DNA is
responsible for coding most of the genetic information
in an organism and is expressed in an organisms
physical appearance, personality, and behavior. With
the enormous amount of information that completes an
organisms genetic make up, it is astounding to
discover how it all fits within the DNA molecule. The
answer lies within the DNAs double-stranded helix
structure.
The basic units of DNA are nucleotides.
Nucleotides are made up of three covalently bonded
parts, a nitrogenous base (purines: adenine and
guanine, pyrimidines: thymine and cytosine), a
deoxyribose sugar, and a phosphoric acid residue. An
N-glycosidic bond links the C-1 carbon of the
deoxyribose sugar to the N-9 nitrogen in purines or the
N-1 nitrogen in pyrimidines.
APPENDIX
Weight centrifuge tube: 6.68g
Weight centrifuge with sample: a.) 7.02g
Weight sample: a.) 0.34g b.) 0.31g
b.) 6.98
Conc.
(10%)(x)=(1%)(10mL+x)
x=0.909mL
x100= 3.4% w/v
dsDNA concentration=50
x dilution
factor
=50
x0.6598x
= 3.299