• Journal of Microbiology
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• 제34권3호
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• pp.229-235
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• 1996

Microgram quantities of DNA per gram soil were recovered with SDS- based and freeze-and thaw procedures. The average DNA fragment size was > 23 Kb. This method generated minimal shearing of extracted DNA. However, the DNA extracts still contained considerable amounts of humic impurities sufficient to inhibit PCR. Several approaches were used to reduce the interferences with the PCR (use of CTAF in extraction step, Elutip-d column purification, addition of BSA to PCR buffer) to accomplish PCR with DNA extract as a template. Most of the DNA extracts were not digested completely by restriction endonuclease, and CTAB-TREATED ane Elutip-d column purified DNA extracts were partially digested. Regarding as restriction enzyme digestion, all PCRs failed to amplify 16S rRNA gene fragments in the DNA extracts. In the case of DNA extracts only where BSA was added to PCR buffer, PCR was successfully conducted whether the DNA extracts were treated with CTAB or purified with columns. However, these two treatments were indispensable for humic impurity-rich DNA extracts to generate the PCR-compatible DNA samples. Direct extraction of DNA, coupled with these procedures to remove and relieve interferences by humic impurities and followed by the PCR, can be rapid and simple method for molecular microbiological study on soil microorganisms.

• 한국환경농학회지
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• 제30권4호
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• pp.367-376
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• 2011

논 토양의 미생물 생태 다양성을 조사하기 위한 효과적인 방법으로 qRT-PCR을 적용하고자 본 연구를 수행하였다. 논 토양 미생물의 gDNA를 분리하기 위하여 Mo Bio kit를 사용한 효과적이고 안정적인 gDNA 분리 방법을 확립하였다. 논 토양 미생물 다양성을 qRT-PCR로 검출하기 위하여 bacteria를 세분한 ${\alpha}$-Proteobacteria, ${\beta}$-Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes 다섯 가지 문과 전체 bacteria, 전체 fungi를 구분할 수 있는 특이 primer set을 선정하여 다양한 조건의 시험을 통하여 최종 조건을 확립하였으며 재현성 실험을 통하여 방법의 유의성을 검증하였다.

• 한국토양비료학회지
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• 제37권4호
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• pp.251-258
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• 2004

밭작물에 대한 규산질 비료의 시용이 널리 이루어지고 있으나 적정시용수준이 밝혀져 있지 못하며 또한 밭토양에서의 유효규산 측정방법이 구명되어있지 못한 실정이다. 본 연구는 참외 시설재배지 토양에 대하여 유효규산 측정방법을 구명하기 위해 수행되었다. 경북 성주지역의 참외 시설재배지,10개소의 토양과 참외 잎 시료를 채취하여 가용성 토양 규산 함량과 잎 중의 총 규산 함량을 분석하였다. 가용성 토양 규산은 0.5 N HCI, 1 N sodium acetate buffer (pH 4.0). citric acid 1%, water, Tris buffer (pH 7.0), 그리고 1주간 담수하는 방법 등으로 추출하였으며, 식물체 규산은 autoclave 방법으로 추출하였다. 추출액중의 가용성 규산은 비색법으로 정량하였다. 각 추출방법별로 가용성 토양규산 함량과 식물체규산 함량과의 관계를 비교하였는데, 1 N sodium acetate buffer 방법이 토양 규산과 식물체 규산 관계를 가장 뚜렷한 포화곡선으로 나타내었다. 포화곡선으로부터 산출된 참외 잎 중의 포화 규산 함량은 약 $14g\;SiO_2\;kg^{-1}$이었고 1 N sodium acetate buffer 방법으로 추출할 경우토양 규산 함량 $120mg\;SiO_2\;kg^{-1}$이 참외에 적정한 수준인 것으로 나타났다. 특히 1 N sodium acetate buffer방법의 경우 참외 잎 중의 규산 함량이 포화되는 수준 이하의 토양 규산 함량 범위에서는 토양 규산 함량과 식물체 규산 함량 사이에 유의성 있는 상관관계가 있었다. 따라서 현재 우리나라에서 논토양의 유효규산 측정방법으로 널리 사용퇴고 있는 1 N sodium acetate buffer를 이용한 유효규산 추출방법이 밭토양에도 적용될 수 있을 것으로 판단되나 앞으로 다양한 작물과 토양을 대상으로 계속적인 연구가 요구된다.

• Journal of Magnetics
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• 제10권3호
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• pp.89-92
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• 2005

The analytic possibility of mobile lead contained in soil has been studied using carboxylated magnetic beads. Extraction of heavy metal was performed to contaminated soil that has been collected and supplied for tests. As experiment materials, soil sample, distilled water and magnetic beads were only used. It means that the lead was extracted under neutral condition. In this condition, only the mobile fraction of lead could be extracted by magnetic beads. The mobile lead in the soil was quickly combined with magnetic beads in the mixture process. Then, the magnetic beads were dissolved into acids after collection by external magnetic force, and the lead combined with the beads was eluted and analyzed by Graphite Furnace Atomic Absorption Spectroscopy (GFAAS). In the results of extraction experiments for 3 sandy soils, the efficiency using beads was similar to or higher than that of EDTA (Ethylendiamintetraacetic acid), which is normally used for analyzing mobile heavy metal concentration in soil. With this, it was shown that this method is a more accurate and simple method to analyze mobile lead when analyzing mobile heavy metal concentration in sandy soil, rather than conventional method using EDTA.

• Journal of Radiation Protection and Research
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• 제21권1호
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• pp.9-16
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• 1996

환경시료 중 토양에 대한 방사성 스트론튬인 $^{89}Sr$와 $^{90}Sr$의 분석을 위해 Ca과 Sr을 분리하기 위한 방법으로 Crown ether를 이용한 추출방법을 사용하여 기존의 발연질산법과 비교하여 보았다. Crown ether를 이용한 추출방법을 기존의 발연질산법과 비교하여 보았을 때 높은 Sr 수율을 보였을 뿐만 아니라 분석에 소요되는 시간과 절차가 훨씬 빠르고 간단했다. Crown ether을 이용한 추출방법으로 원전주변 토양에 대한 방사성 스트론튬의 분석을 수행하였으며, 방사성핵종 분석에 의한 환경감시의 효율적인 수행을 위한 스트른튬 분석법을 제시하였다

• 한국지하수토양환경학회지:지하수토양환경
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• 제15권3호
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• pp.1-6
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• 2010

This study investigated the change of metal contamination levels according to amendment of enforcement regulation of the Soil Environmental Conservation Act in Korea. As an analytical result of 87 samples in abandoned metallic mine area, the extracted amount of As, Pb and Cu with aqua regia was 4.3~29.6 times higher than that with hydrochloric acid extraction and the number of samples, which contamination levels were found to exceed soil contamination standards, was also increased. On the other hand, in case of Cd, Zn, and Ni, the number of samples, which contamination levels were found to exceed soil contamination standards, was decreased or similar. These results can be used as a preliminary material in comparison between the soil pollution data accumulated previously and the data obtained by the revised standard method for the examination of soil pollution.

• 한국농공학회:학술대회논문집
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• 한국농공학회 1999년도 Proceedings of the 1999 Annual Conference The Korean Society of Agricutural Engineers
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• pp.447-453
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• 1999

Salt accumulation is one of themajor problems in soils under protected cultivation . Since protected cultivation does not have rainfall or excessive irrigation, salt accumulaiton inthe soils is inevitable. In this study, characteristics of salt accumulation in soil column were ivestigated, and a salt-extracting method was tested to see its effectiveness. The results showed that the concentration of salt in top soil layers increased and electrical conductivity as thesalt concentration decreased especially in the top soil layer .When extraction medium was applied.

• Journal of Applied Biological Chemistry
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• 제56권2호
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• pp.95-100
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• 2013

현재 토양 생태에서 토양미생물은 유기물 분해, 질소 순환, 식물의 질소 이용 등 중요한 역할을 하고 있어, 토양 내 미생물 다양성을 분석하기 위한 연구는 지속적으로 진행되어 오고 있다. 본 연구에서는 논 토양의 미생물 생태 다양성을 조사하기 위한 효과적인 방법으로 denaturing gradient gel electrophoresis (DGGE)를 적용하고자 본 연구를 수행하였다. 논 토양 미생물의 DNA를 분리하기 위하여 lysis buffer method, skim milk bead method, sodium phosphate buffer method, Epicentre SoilMaster DNA extraction kit (Epicentre, USA), Mo Bio PowerSoil kit (Mo Bio, USA)를 이용하여 토양 내 gDNA 최적 추출방법을 확인하였다. 그 결과 Mo Bio PowerSoil kit를 사용하였을 때 Shannon 다양성지수가 세균 3.3870, 진균 3.6254으로 미생물 다양성 분석시에 가장 효과적이었다. DGGE 분석을 위한 조건은 세균의 경우 6% polyacylamide gel, 45-60% denaturing gradient였고, 진균의 경우 6% polyacrylamide gel, 45-80% denaturing gradient에서 최적 분석조건을 보였다. 위의 분석법을 적용하여 논 토양내의 미생물 군집의 변화를 살펴보면 시간의 변화 요인에 의해 미생물 변화가 일어나는 것을 알 수 있었다. 본 연구에서 사용된 DGGE 분석법을 통해 논토양 미생물의 분석 가능성을 제시 할 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제26권12호
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• pp.2051-2059
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• 2016

To date, there has been no employment of a magnetoelastic (ME) biosensor method to detect Salmonella enterica serovar Typhimurium in soil. The ME biosensor method needs to be investigated and modified for its successful performance. The filtration method, cation-exchange resin method, and combinations of both methods were employed for the extraction of S. Typhimurium from soil. The number of S. Typhimurium and the resonant frequency shift of the ME sensor were then compared using a brilliant green sulfa agar plate and an HP 8751A network analyzer. A blocking study was performed using bovine serum albumin (BSA), polyethylene glycol (PEG), and casein powder suspension. Finally, the modified ME biosensor method was performed to detect S. Typhimurium in soil. The number of S. Typhimurium was significantly decreased from 7.10 log CFU/soil to 4.45-4.72 log CFU/soil after introduction of the cation-exchange resin method. The greatest resonant frequency shift of the measurement sensor was found when employing centrifugation and filtration procedures. The resonant frequency shift of the PEG-blocked measurement sensor was $3,219{\pm}755Hz$, which was significantly greater than those of the BSA- and casein-blocked ME sensor. The optimum concentration of PEG was determined to be 1.0 mg/ml after considering the resonant shift and economic issue. Finally, the modified ME biosensor method was able to detect S. Typhimurium in soil in a dose-response manner. Although these modifications of the ME biosensor method sacrificed some advantages, such as cost, time effectiveness, and operator friendliness, this study demonstrated a novel approach of the ME biosensor method to detect S. Typhimurium in soil.

• 한국지하수토양환경학회지:지하수토양환경
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• 제23권2호
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• pp.15-22
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• 2018

Aqua regia extraction for the quantification of heavy metals and As in contaminated soils was investigated as one of preliminary steps for on-site monitoring using sensor technology. Soil samples were taken from closed railway depot, closed mines, and closed refinery and various extraction conditions including $30^{\circ}C/15min$, $30^{\circ}C/30min$, $30^{\circ}C/60min$, $30^{\circ}C/120min$, $80^{\circ}C/15min$, $80^{\circ}C/30min$, $80^{\circ}C/60min$, $80^{\circ}C/120min$ were tested. The optimal extraction condition was determined as $80^{\circ}C/60min$ because the extraction efficiencies of Zn were relatively low and did not reach the targeted level (80-100% of original concentrations) for $30^{\circ}C$ conditions. It was found that the fractionation of heavy metals and As using the sequential extraction method was useful to understand the degree of metal extraction. In order to enhance the extraction efficiency within short extraction time, ultrasound technology using a 20 kHz horn-type sonicator was additionally used for $30^{\circ}C/15min$. It was revealed that ultrasound could significantly enhance the extraction efficiency and pulsed irradiation showed higher efficiency than continuous irradiation due to the less formation of bubble clouds. However high temperature condition ($80^{\circ}C$) was required to achieve high extraction efficiency for Zn in spite of the use of ultrasound.