DNA ISOLATION ON CAPTIVE SUMATRAN ELEPHANT IN ELEPHANT TRAINING CENTER, WAY KAMBAS NATIONAL PARK: A FIRST STEP TOWARDS ITS ID CARD
TOWARDS ITS ID CARD
E l l y L . R u s t i a t i 1 * , P r i y a m b o d o 1 , S i t i A s i y a h 1 , D e d i C a n d r a 2 , D i a h E . A n g g r a i n i 2 , E l i z a b e t h D . K r i s m u n i a r t i 2 , E k o A . S r i h a n to 3 , L i z a A n g e l i y a 3 , N u n i n g N u r c a hy a n i 1 , E n n y S a s w i ya n t i 3 DNA ISOLATION ON CAPTIVE SUMATRAN ELEPHANT IN ELEPHANT TRAINING CENTER, WAY KAMBAS NATIONAL PARK: A FIRST STEP
1 D e p a r t m e n t o f B i o l o g y , F a c u l t y o f M a t h e m a t i c s a n d N a t u r a l S c i e n c e s , U n i v e r s i t y o f L a m p u n g 2 E l e p h a n t T r a i n i n g C e n t e r , W a y K a m b a s N a t i o n a l P a r k , E a s t L a m p u n g 3 V e t e r i n a r y B u r e a u , L a m p u n g P r o v i n c e , B a n d a r L a m p u n g
ABSTRACT
E le ph an t Tr aini ng Ce nter ( E TC ) Way Kam b a s N ati o nal Par k (W K NP ) wa s built
to s u pp or t h um a n -ele ph a nt m itig ati o n c onfli ct . T he sm all p o p ulatio n o f
c a pti ve s um atr an ele pha nt in ET C W K NP n ee d a c om pr e he n siv e str ateg y i n
o r d er to m ain tai n th e ge neti c v ar iati o n of e a c h i ndiv i du al an d av oid
i nbr e edi n g dr ive . C ur r en tl y, ge neti c st u die s h ave o pe ne d n ew fi eld st u die s in
e c ol o g y, i n clu de d c o n ser va tio n e c ol og y. Patter ns i n v ar iati o n of p o p ula ti on
h a s be en i nv e stig ate d by m ol e cul ar m eth o d su pp or ti ng s pe ci es c on ser va ti on
e f for t . T h e c a ptiv e s um a tr a n elephant’s ID Card is a necessar y in databaseb uil din g , w hi c h in cl ud ed m or p h ol og y, he alth s t atu s , a nd ge neti c pro file.
G e neti c pr ofile i n ea c h I D C ar d was fille d by c y tog eneti c an d m ol ec u lar
pr ofile for R A DP r e sult , th at i nitiated with DNA i s olati o n. T he DN A s our ce s
c o lle cted by bl o o d s am p lin g pr oto c ol de s cr ib ed by A si ya h e t al . ( 2016 ) fr om
c a pti ve s umatr an ele pha nt in ETC , W K NP, an d be car r ie d to la b or ator y in c old
oc o n diti o n. T he D NA s o urc e s stor ed at 4 C a n d iso l ate d foll owin g c om m er ci al
opr oto c ol . T h e r e s ult o f D N A i s olati o n stor e d a t - 20 C u ntil am plifi ca ti on
a n al y sis . D NA i s ola tio n wa s su c c es s full y d on e , for f ur ther in div i du al gen etic
I D b u i l d ing.Ke y wo r d s: C o n s e r va tio n , D N A i s o l a t io n, s u m a tr a n elephant’s ID Card, WKNP
INTRODUCTION
Elephants belongs to the order of Proboscidea
with only two living genera, Loxodonta and Elephas.
The asian elephant population has shown
constant sign of decreased and in 1986 was listed as endangered species by the IUCN .
The lost of habitat combines with high
human-population density, like in Asia, has resulted in Human -Elephant Conflict
To mitigate the conflict, the Elephant Training
Centre (ETC) in Way Kambas National Park , East Lampung, Sumatera, Indonesia was
INTRODUCTION
The small population of captive sumatran
elephant in ETC WKNP need a comprehensive strategy in order to maintain the genetic variation of eac h individual and avoid inbreeding drive.
In WKNP, DNA bank data needs to be built
(Priyambodo et al., 2017) and blood sampling were carried out in collaboration with WKNP and medical teams ( Rustiati et al., 2017).
Genetic studies have become one of new field
studies in ecology, included conser vation ecology.
METHODS This project is conducted under research grant from Directorate of Research and Community Ser vices, Higher Education Indonesia .
The DNA sources collected by whole blood sampling protocol described by Asiyah et al.
(2016) from captive sumatran elephant in ETC,
WKNP. DNA isolation was conducted successfully by silica gel colomn methods with commercial QIAGEN Dneasy and tissue kit for blood and tissue.
Three main steps included cell preparation/lysis,
DNA isolation from contaminants and DNA precipitation .DNA isolation success was qualitatively analyses by 1% agarose gel electrophoresis technique and
RESULTS AND DISCUSSION
RESULTS AND DISCUSSION
Twenty eight individual elephants have been whole blood
sampled.
DNA was successfully separated from protein and other
component (Figure 1) and kept in -40
C. Qualitative test using agarose gel electrophoresis tec hnique.
RESULTS AND DISCUSSION
RESULTS AND DISCUSSION
In electrophoresis process 1% agarose gel was applied as
stationer phase, and Tris-acetate EDTA (TAE) as movement phase.
DNA genome was separated in agarose gel based on electro
mobility of its negative c harge of DNA molecule in cer tain distance.
Loading dye addition on DNA may clarify migration distance.
Sumatran elephant’s DNA genome was predicted more than 10kb. It is shown by its shor t migration distance from the well.
Isolated DNA will be used for fur ther molecular analysis, PCR-
RAPD and DNA sequencing.
CONCLUSION
Twenty eight whole blood samples have been successfully
prepared for isolation of DNA . DNA samples were available
o
for fur ther individual genetic ID building, and kept in -20 C.
ACKNOWLEDGEMENT
Our high gratitude to Directorate of Researc h and Community Ser vices, Higher Education Indonesia for researc h grant, Way Kambas National Park for the scientific collaboration and Veterinar y Bureau Lampung Province for Biomolecular analysis works. Our thanks to Dr. Mikael Jazdzyk for literature suppor ts.