Journal
SCIENCE & JUSTICE
Volume 61, Issue 5, Pages 573-578Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.scijus.2021.07.002
Keywords
CTAB; DNA; Extraction; PCR Amplification; Tropical Timber
Categories
Funding
- Ministry of Research and Technology/National Agency for Research and Innovation (RISTEK/BRIN) of Republic of Indonesia [1/E1/KP.PTNBH/2021, 1918/IT3.L1/PM/2021]
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This study compared the DNA isolation results of various dried and fresh wood samples using two extraction methods (CTAB and Qiagen DNeasy Plant Mini Kit). While dried wood samples showed poor visualization, fresh wood samples had higher DNA concentration in CTAB-extracted samples compared to QDPMK-extracted samples.
The successful isolation of DNA (Deoxyribonucleic Acid) is essential for the investigation process of forestry molecular genetics. Samples used are usually retrieved either from soft or juvenile plant organs because of their excellent DNA source. However, in certain cases, aforesaid samples are hard to obtain, as for forensic purposes. Alternatively, woods possess potential as alternative source of DNA whose extraction method has been developed with varying degrees of success. However, to date, effectiveness on tropical wood grown in Indonesia has not been widely reported. Therefore, objective of this study was to compare the results of DNA isolation of various dried and fresh wood samples by using two isolation methods: Cetyl Trimethyl Ammonium Bromide (CTAB) and Qiagen DNeasy Plant Mini Kit (QDPMK). Extraction results were visualized in agarose gels and quantified using Nanophotometer NP80 Implen which were then amplified using two universal primers: ITS and rbcL for detecting DNA signals. Extraction results from dried wood indicated no visualization in the gel, while fresh wood samples showed thick smeared bands on both extraction methods. Quantity test results denoted higher concentration in CTAB-extracted samples compared to samples extracted using QDPMK, in both types of samples, even though both resulted in optical density ratios outside the range of purity (lambda 260/280: 1,8-2,0 and lambda 260/230: 2,0, respectively). Success rates of ITS and rbcL primary amplification in dried wood samples were quite low yet outputs from the two methods did not differ significantly. Meanwhile, outcome of ITS and rbcL amplification on fresh wood samples had a fairly high success.
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