MOLECULAR SEXING OF THE SAMBAR DEER

(RUSA UNICOLOR) USING PCR

AMPLIFICATION OF THE AMELOGENIN GENE
AND DBY GENE

Author: LaTanya Downer
Jothi Malar Panandam, Kamariah
Jamhari
INTRODUCTION
The Sambar is the largest deer native of the
Indian subcontinent, southern China, and
Southeast Asia.
Its scientifically referred to as Rusa unicolor.
Polymerase chain reaction (PCR) is used in molecular
biology to amplify a single or a few copies of DNA and
create millions of copies.
Sequencing is the process of determining the precise
order of nucleotides within a DNA molecule.
Molecular sexing is not only an attractive means to
differentiate the sexes of sexually monomorphic species,
but may be used to identify the gender of the killed
animal from the meat and body fluid, as well as to
identify the species.
PROBLEM STATEMENT
There is little genetic information about Sambar
deer.
I want to used this research opportunity to add
information about the deer using molecular sexing.

OBJECTIVES
To learn some techniques in Molecular Biology.
To determine the amplification products and nucleotide
sequences of the X and Y chromosomes linked
Amelogenin genes (AMELX and AMELY) and the Y-
linked DEAD-box RNA helicase Y gene (DBY) in the
Sambar deer (Rusa unicolor).
To compare the nucleotide sequences of AMELX and
AMELY between the male and female Sambar deer.


METHODS & MATERIALS
DNA EXTRACTION
Blood
Sample
DNeasy Blood & Tissue
Kit
Biophotometer
Centrifuge
Used in the DNA
extraction
Used to check
concentration of DNA
Thermal Cycler
POLYMERASE CHAIN REACTION (PCR)
Used to amplify
specific region of the
DNA.
Amelogenin Region
and DBY Region
Specific Protocol
The DEAD-Box RNA Helicase Y (DBY / dby) region in Y chromosome
of human and mouse.

Source: Mazeyrat et al., 1998
PRIMER SE 47 & 48 IS DESIGN BASE ON THE
BOVINE AMELOGENIN LOCUS
ELECTROPHORESIS
SE 47 & 48 uses
Metaphor Agarose gel
(4%)
DBY 8 uses Agarose
gel Electrophoresis
(1.6%)

Gel Documentation System
GEL VIEWING
Test product for electrophoresis
PURIFICATION
Expin
Tm
Combo GP

RESULTS

SE 47&48 Metaphor
Agarose Gel
DBY 8 Agarose Gel
SEQUENCING
BioEdit is a program used to view the nucleotide
sequence of the PCR amplified and sequenced
DNA fragments.

ClustalW2 is a general purpose multiple sequence
alignment program for DNA or proteins. It is used to
compare the sequences of PCR amplicons of
different individuals and species, and in this case
two sexes, i.e. of AMELX and AMELY.

CONCLUSION
Primer SE 47 & 48 and DBY 8 can detect the sex of
the animal.



ACKNOWLEDGEMENT
Jothi Malar Panandam]

Kamariah Jamhari