EKSTRAKSI DNA DARI HERBARIUM ANGGREK

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