EKSTRAKSI DNA DARI HERBARIUM ANGGREK

Abstract

DNA extraction is the first step to study plant systematic and biodiversity analysis using
molecular markers. This study aimed to conduct DNA extraction from herbarium materials
using different extraction methods. A total of 0.05 grams of herbarium powders of
Calantheemarginata (Blume) Lindl. and Goodyera procera(Ker-Gawl) Hook. (terrestrial
orchid) were used for samples by three different methods. The first method was from Doyle
and Doyle with modification of incubation time for 1,5 hours at 65oC and increasing EDTA
concentration to 50 mM. Second method was Dellaporta et al. with modification of incubation
time for 1,5 hours (at 65oC) and increasing EDTA concentration to 100 mM. Third method
was Rogers and Bendich with modification of incubation time for 1,5 hours (65oC) and
adding ethanol twice. The results of electrophoresis revealed that method of Doyle and Doyle
obtained DNA from C. emarginata herbarium, while method from Rogers and Bendich,
unfortunately it was inconsistent. The method from Dellaporta et al.obtained DNA from G.
procera herbarium, while method from Doyle and Doyle revealed inconsistent DNA forG.
procera. PCR-RAPD revealed the quality of DNA isolated using Doyle and Doyle method
was not optimal, showed by unclear patterns of DNA bands. PCR-RAPD using DNA isolated
with method from Rogers and Bendich revealed clearer DNA bands but only for small size
fragment.
Keywords : orchid, DNA extraction, herbarium, PCR