82 156 1 SM
82 156 1 SM
ABSTRACT
Potential identification of endophytic microorganisms can be done through genetic
analysis and gene diversity analysis of bacteria. Extracting high-purity DNA directly
from roots has become essential for the study of microorganisms in environmental
samples. This study aims to perform DNA isolation of endophytic bacterial roots from
Vetiveria zizanioides, L. without cultivation stages. In the stages of DNA isolation
directly is required several steps including surface sterilization, extraction and
determination of the quality and quantity of the result. Direct DNA extraction of
endophytic bacterial roots from Vetiveria zizanioides L, has been successfully
performed with the purity 1.798 and concentration reached 780 g / ml.
Key Word: DNA extraction, Endophytic, Vetiveria zizanioides
METODE
Pengambilan sampel. Isolasi mikroba
endofit dilakukan dengan mengambil
sampel akar tanaman Vetiveria zizanioides
L.,
kemudian
dilakukan
sterilisasi
permukaan menggunakan larutan etanol
75% selama 1 menit, bayclin 25% selama 5
menit, dan terakhir dicuci dengan etanol
75% selama 30 detik. Setelah disterilisasi
permukaan, akar kemudian dipotong lebih
halus dan dimasukan pada cairan fisiologis
NaCl 0,98% pada tabung reaksi, kemudian
divorteks selama 1,5 jam (Lumyong et al.,
2001).
Isolasi DNA. Isolasi DNA dilakukan
dengan mengambil 3 ml hasil vorteks pada
tabung 1,5 ml, kemudian proses isolasi
DNA
dilakukan
menggunakan
EG
1
EG
2
1.
Hasil
kuantifikasi
secara
2.
Kode
EG1
EG2
Absorbansi
[DNA
Purity
(nm)
260 280
0,1 0,08
(A260/
g/ml
780
280)
1,798
340
1,625
56
0,0
0,04
68
DAFTAR PUSTAKA
Moffitt,
Sessitsch, A.,
Reiter,B., Pfeifer,U.,
Wilhelm,E. (2002). Cultivationindependent population analysis
of bacterial endophytes in three
potato
varieties
based
on
eubacterial and Actinomycetesspecies PCR of 16S rRNA genes.
FEMS Microbiology Ecology. 39,
23-32.
Streit, W. R., & Schmitz, R. A. (2004).
Metagenomicsthe key to the
uncultured microbes. Current
Opinion in Microbiology. 7, (5),
492-498.
Strobel,
G.,
Daisy,
B.
(2003).
Bioprospecting for Microbial
Endophytes and Their Natural
Products. Microbiology and
Molecular Biology Reviews. p.
491502.
email: fauziedu247@gmail.com