• Mycobiology
• /
• 제42권4호
• /
• pp.311-316
• /
• 2014

Lichen studies, including biodiversity, phylogenetic relationships, and conservation concerns require definitive species identification, however many lichens can be challenging to identify at the species level. Molecular techniques have shown efficacy in discriminating among lichen taxa, however, obtaining genomic DNA from herbarium and fresh lichen thalli by conventional methods has been difficult, because lichens contain high proteins, polysaccharides, and other complex compounds in their cell walls. Here we report a rapid, easy, and inexpensive protocol for extracting PCR-quality DNA from various lichen species. This method involves the following two steps: first, cell breakage using a beadbeater; and second, extraction, isolation, and precipitation of genomic DNA. The procedure requires approximately 10 mg of lichen thalli and can be completed within 20 min. The obtained DNAs were of sufficient quality and quantity to amplify the internal transcribed spacer region from the fungal and algal lichen components, as well as to sequence the amplified products. In addition, 26 different lichen taxa were tested, resulting in successful PCR products. The results of this study validated the experimental protocols, and clearly demonstrated the efficacy and value of our KCl extraction method applied in the fungal and algal samples.

• 한국환경과학회지
• /
• 제27권12호
• /
• pp.1205-1214
• /
• 2018

During the past few decades, significant increase in the consumption of coffee has led to rapid increase in the production of coffee waste in South Korea. Spent coffee waste is often treated as a general waste and is directly disposed without the necessary treatment. Spent Coffee Grounds (SCGs) can release several organic contaminants, including caffeine. In this study, leaching tests were conducted for SCGs and oxidative degradation of caffeine were also conducted. The tested SCGs contained approximately 4.4 mg caffeine per gram of coffee waste. Results from the leaching tests show that approximately 90% of the caffeine can be extracted at each step during sequential extraction. Advanced oxidation methods for the degradation of caffeine, such as $UV/H_2O_2$, photo-Fenton reaction, and $UV/O_3$, were tested. UV radiation has a limited effect on the degradation of caffeine. In particular, UV-A and UV-B radiations present in sunlight cause marginal degradation, thereby indicating that natural degradation of caffeine is minimal. However, $O_3$ can cause rapid degradation of caffeine, and the values of pseudo-first order rate constants were found to be ranging from $0.817min^{-1}$ to $1.506min^{-1}$ when the ozone generation rate was $37.1g/m^3$. Additionally, the degradation rate of caffeine is dependent on the wavelength of irradiation.

• Archives of Pharmacal Research
• /
• 제17권3호
• /
• pp.175-181
• /
• 1994

The solid-phase extraction (SPF) with subsequent tert-butyldimethylsilyl (TBDMS) derivatization was investigated for the rapid profiling and screening of various carboxylated non-steroidal antiinflammatory drugs (NSAIDs) simultaneously in biological fluid samples. Compared to the conventional SPF in adsorption mode using Chromosorb 102, Chromosorb 107, Carbopak B and Thermosorb, the SPF in partition mode using Chromosorb P as the adsorbent, and ethyl acetate/methylene chloride as the eluting solvents provided hightest overall recovenies of the NSAIDs from aqueous solutions with good precision. The solid-phase extracted NASIDs were silylated with N-methyl-N-(tert-butyldimethylsily)trifuoroacetamide to TBDMS derivatives and directly analyzed by capillary gas chromatography and gs chromatography-mass spectrometry. The usefulness of the present method was examined for the profilling and screening of saliva, serum and urine samples for various NSAIDs simultaneously.

• 대한약학회:학술대회논문집
• /
• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
• /
• pp.118.3-119
• /
• 2003

Diazepam (DZ) is one of the most frequently prescribed drugs as an antianxiety agent, muscle relaxant, and anticovulsant and sometimes causes intoxication due to accidental overdose, misuse or abuse. Screening or confirmation methods for DZ and NDZ in plasma are very important for clinical and toxicological studies and in forensic cases. GC/MS assay with SPE was developed for the determination of diazepam and its metabolite, nordiazepam in human plasma. Diazepam in plasma was extracted by a rapid and sensitive procedure based on C18 bonded-phase extraction. (omitted)

• 식물병연구
• /
• 제10권1호
• /
• pp.53-57
• /
• 2004

CGMMV는 한국에서 수박의 주요 병원균이고, 수박 생산에 심각한 영향을 미친다. 이 연구에서는 박 종자의 CGMMV를 RT-PCR을 이용하여 신속하고 민감하게 검정하는 진단방법을 개발하였다. CGMMV-W의 외피 단백질 유전자 sequence에서 제작된 CGMMV에 특이적인 primer인 Wmfl과 Wmrl은 RT-PCR에 의해 420 bp의 증폭산물을 증폭하였다. RT-PCR에 의한 진단을 위하여 바이러스 추출과정을 간소화하고 종자 추출물의 반응 억제물질을 감소시키기 위해 ethanol 침전, double filtration, PEG 침전, phenol/chloroform/isoamyl alcohol에 의한 추출법을 비교하였으며 phenol/chloroform/isoamyl alcohol에 의 한 추출법이 민감성이 강한 방법으로 선발되었다. RT-PCR을 위해 선발된 primer들과 추출법은 1,000립의 건전 종자에 1립의 이병 종자를 혼합한 수준까지 판별이 가능하였다. 신속하고 민감한 RT-PCR에 의한 본 검정방법은 높은 반응 억제물질을 함유하는 박 종자에서 CGMMV의 특이적인 진단을 위해 유용한 방법이다.

• Plant Biotechnology Reports
• /
• 제4권1호
• /
• pp.49-52
• /
• 2010

A DNA extraction method using Chelex 100 is widely used for bacteria, Chlamydomonas, and animal cell lines, but only rarely for plant materials due to the need for additional time-consuming and tedious steps. We have modified the Chelex 100 protocol and successfully developed a rapid and simple method of DNA extraction for efficient PCR-based detection of transgenes from a variety of transgenic plant and algal species. Our protocol consists of homogenizing plant tissue with a pestle, boiling the homogenized tissue in a microfuge tube with 5% Chelex 100 for 5 min, and centrifuging the boiled mixture. The supernatant, which is used for PCR analysis, was able to successfully amplify transgenes in transgenic tobacco, tomato, potato, Arabidopsis, rice, strawberry, Spirodela polyrhiza, Chlamydomonas, and Porphyra tenera. The entire DNA extraction procedure requires <15 min and is therefore comparable to that used for bacteria, Chlamydomonas, and animal cell lines.

• Journal of Microbiology and Biotechnology
• /
• 제15권3호
• /
• pp.595-602
• /
• 2005

A one-step real-time quantitative RT-PCR assay in combination with automated RNA extraction was evaluated for routine testing of HCV RNA in the laboratory. Specific primers and probes were developed to detect 302 bp on 5'-UTR of HCV RNA. The assay was able to quantitate a dynamic linear range of $10^7-10^1$ HCV RNA copies/reaction ($R^2=0.997$). The synthetic HCV RNA standard of $1.84{\pm}0.1\;(mean{\pm}SD)$ copies developed in this study corresponded to 1 international unit (IU) of WHO International Standard for HCV RNA (96/790 I). The detection limit of the assay was 3 RNA copies/reaction (81 IU/ml) in plasma samples. The assay was comparable to the Amplicor HCV Monitor (Monitor) assay with correlation coefficient r=0.985, but was more sensitive than the Monitor assay. The assay could be completed within 3 h from RNA extraction to detection and data analysis for up to 32 samples. It allowed rapid RNA extraction, detection, and quantitation of HCV RNA in plasma samples. The method provided sufficient sensitivity and reproducibility and proved to be fast and labor-saving, so that it was suitable for high throughput HCV RNA test.

• 대한생식의학회:학술대회논문집
• /
• 대한불임학회 1999년도 제38차 추계 학술대회
• /
• pp.73-74
• /
• 1999

• 한국환경보건학회지
• /
• 제21권3호
• /
• pp.87-95
• /
• 1995

A rapid and selective co-extraction systems of copper and zinc-thiocyanate complex into various types of alkylamine for the simultaneous determination of two metal ions by atomic absorption spectrometry and ion chromatograph have been proposed. The quantitative extractions of Cu(II) and Zn(II) at 0.1 M-thiocyanate and 0.1 M-HCI were achieved with Aliquat 336-$CHCl_3$. The detection limits of Cu and Zn were 2 ppb and 0.9 ppb respectively.

• 전력전자학회:학술대회논문집
• /
• 전력전자학회 2001년도 Proceedings ICPE 01 2001 International Conference on Power Electronics
• /
• pp.650-655
• /
• 2001

Wind speeds can vary rapidly and wind turbines cannot easily follow these variations because of their inertia and aerodynamic characteristics. For maximum energy extraction. the turbine blades should operate at their optimum tip speed ratio, but with rapid changes in wind speed. this is usually not possible. To improve the energy extraction from turbulent wind, it is necessary to establish an effective measure of the high frequency component of the wind. and then to use this measure to optimise the operation of the turbine controller for maximum energy extraction. This paper presents an approach for combining readings from three anemometers into a composite wind speed measurement. and using this signal to control the operation of a permanent magnet generator to achieve maximum energy extraction. The method combines simulation and experimental investigations into a heuristic algorithm. and demonstrates its effectiveness with field trials.