UUL SHOVI NURKAMILA
Jurusan Biologi F.MIPA Universitas Udayana, Kampus Bukit Jimbaran – Bali

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EKSTRAKSI DNA DARI HERBARIUM ANGGREK NURKAMILA, UUL SHOVI; PHARMAWATI, MADE
SIMBIOSIS Journal of Biological Sciences Vol II, No 1, Tahun 2014
Publisher : SIMBIOSIS Journal of Biological Sciences

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Abstract

DNA extraction is the first step to study plant systematic and biodiversity analysis usingmolecular markers. This study aimed to conduct DNA extraction from herbarium materialsusing different extraction methods. A total of 0.05 grams of herbarium powders ofCalantheemarginata (Blume) Lindl. and Goodyera procera(Ker-Gawl) Hook. (terrestrialorchid) were used for samples by three different methods. The first method was from Doyleand Doyle with modification of incubation time for 1,5 hours at 65oC and increasing EDTAconcentration to 50 mM. Second method was Dellaporta et al. with modification of incubationtime for 1,5 hours (at 65oC) and increasing EDTA concentration to 100 mM. Third methodwas Rogers and Bendich with modification of incubation time for 1,5 hours (65oC) andadding ethanol twice. The results of electrophoresis revealed that method of Doyle and Doyleobtained 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 methodwas not optimal, showed by unclear patterns of DNA bands. PCR-RAPD using DNA isolatedwith method from Rogers and Bendich revealed clearer DNA bands but only for small sizefragment.Keywords : orchid, DNA extraction, herbarium, PCR