• Korean Journal of Breeding Science
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• v.40 no.3
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• pp.286-290
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• 2008

Many important traits have been tagged allowing plant breeders to apply marker assisted selection (MAS) in rice. PCR itself is simple to set up, and requires little hands-on time. However, a crucial limiting step of MAS programs is the reliable and efficient extraction of DNA which can be performed on thousands of individuals. In this study, We describe a modification of the DNA extraction method, in which cetyltrimethylammonium bromide (CTAB) is used to extract DNA from leaf tissues for suitable MAS in rice. We followed the standard 2% CTAB extraction method in all the procedure. In addition we used the 1.2 ml 8-strip tube instead of 1.5 ml E-tubes to fit the 8-multichannel pipette and employ the 96 well plate to use the swing bucket centrifuge. Our modified CTAB DNA extraction method offers several advantages with respect to traditional and simple methods. 1) adult leaf samples collected in paddy field are applicable. 2) 96 leaf samples can be homogenized only one-time by using tungsten carbonate bead and 96well block. 3) semiautomatic loading method using 8-multichannel pipette from DNA extraction to electrophoresis of PCR products. 4) our system can extract about 400 leaf samples per day by only one technicion. Therefore, this method could be useful for marker assisted breeding in rice.

• Analytical Science and Technology
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• v.21 no.4
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• pp.338-343
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• 2008

Extraction of DNA from skeletal material is of great importance in the identification of human remains, but is particularly difficult because the high amount of microbial DNA was often co-extracted with human bone DNA. We found that a phenol/chloroform extraction, followed by ultrafiltration, and cleanup by via the $QIAquick^{(R)}$ PCR purification kit yields higher amounts of human genomic DNA compared with extraction by the column affinity $method^{(R)}$ alone. Ultrafiltration extraction of human DNA from ten exhumed bone samples yielded $0.041-1.120ng/{\mu}L$ DNA (mean = $0.498ng/{\mu}L$ DNA), and purification using the column affinity resulted in $0.016-0.064ng/{\mu}L$ DNA (mean = $0.034ng/{\mu}L$ DNA). Although the STR genotyping by the column affinity method was partially successful, all DNA samples by the ultrafiltration method produced full profiles from the multiplex PCR. The efficiency of STR genotyping was in accordance with the amounts of the human DNA extracted.

• Korean Journal of Veterinary Research
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• v.54 no.1
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• pp.55-57
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• 2014

Paratuberculosis caused by Mycobacterium avium subsp. paratuberculosis (MAP) has extended latent periods of infection. Due to this property, difficulties in the detection of fecal shedder have been raised. A newly designed method for DNA extraction from fecal specimens, mGITC/SC was evaluated in terms of diagnostic efficiency. The detection limit of IS900 real-time PCR was about 50 MAP (1.5 cfu) in 250 mg of feces (6 cfu per g). Also, this DNA extraction method was faster and cheaper than that using commercial kit or other methods. Consequently, the mGITC/SC is an economical DNA extraction method that could be a useful tool for detecting MAP from fecal specimens.

• Journal of Oral Medicine and Pain
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• v.20 no.2
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• pp.515-528
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• 1995

The human genomic deoxyribonucleic acid(DNA) was extracted from the pulp, dentin of 22 teeth by clelex, phenol methods. Samples of the tooth-derived DNA were amplified by polymerase chain reaction(PCR), electrophosed for sex determination by detection of X-Y homologus amelogenin gene and D1S80 locus detection The following results have been achieved. 1. Chelex and phenol method are effective to sex determination in the pulp and dentin 2. Chelex method is not suitable for detection of D1S80 locus. 3. Concentration and purity of DNA for teeth using chelex method is lower than using phenol method. From the above investigation, chelex method is simple, rapid for sex determination, but it is not suitable for detection of VNTRs.

• ALGAE
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• v.18 no.1
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• pp.65-69
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• 2003

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.spc1
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• pp.269-273
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• 2006

The study of molecular markers to improve crops largely depends on the availability of rapid and of efficient DNA extraction methods. Here we developed a cheap and convenient method to isolate genomic DNA from rice grains suitable for large-scale microsatellite analysis. We confirmed that the isolated rice DNA is suitable for PCR analysis with STS marker and SNP marker, as well as microsatellite marker. Further, we established high-throughput DNA extraction system in a 96-well plate format which make it possible high-throughput analysis of microsatellite markers with rice grains. This implies that the new method could be a useful tool for other types of marker analysis in large scale.

• Korean Journal of Food Science and Technology
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• v.35 no.4
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• pp.591-597
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• 2003

To compare the quality of genomic DNA extracted from potato for PCR detection, four different methods, such as silica-based membrane method, silica-coated bead method, STE solution treatment, and CTAB-phenol/chloroform method, were evaluated. Also, to remove an excessive carbohydrate from the potato, ${\alpha}$- and ${\beta}$-amylase were used individually and in combination. When used both silica-based membrane method and silica-coated bead method combined with enzymes, the genomic DNAs were extracted from the raw potato with high purity for PCR. However, the silica-coated head method combined with enzyme treatment was the most efficient for extraction of the genomic DNA from the frozen fried potatoes. When applied with STE solution, the highly purified DNA was extracted from the raw potatoes without enzyme treatment in adequate yield for PCR. In cases of processed potatoes, such as frozen-fried potato and fabricated potato chips, CTAB-phenol/chloroform method is mostly feasible for DNA extraction and PCR efficacy at high sensitivity. As the results of PCR amplification, 216bp of PCR product was detected on 2% agarose gel electrophoresis, but any amplicons derived from New leaf and New leaf Y gene was not detected in any sample.

• Korean Journal of Veterinary Service
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• v.30 no.2
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• pp.233-241
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• 2007

Tuberculin positive cattle without gross tubercle lesions should be confirmed by the bacteriological examination to determine the state of the infection. To overcome the time-consuming and laborious identification by culture and biochemical tests, polymerase chain reaction (PCR) has been used to identify Mycobacterium bovis. Due to various lipids in the cell wall of Mycobacterium spp, novel methods of DNA extraction from Mycobacterium spp have been developed. In this study, a newly developed guanidium isothiocyanate/silica DNA extraction method was directly applied to specimens from the tuberculin positive cattle. DNAs were directly extracted from the lymph nodes and the major polymorphic tandem repeat (MPTR) and mycobacterial protein of BCG 70 (MPB70) were amplified using PCR. The DNA extraction method using guanidium isothiocyanate/silica was efficient and safe, and the MPTR and MPB70 primers were specific to M bovis. Therefore, MPTR and MPB70 PCRs will be useful for the detection of M bovis in the lymph node from skin-test positive cattle.

• Animal cells and systems
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• v.13 no.3
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• pp.351-356
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• 2009

Ciliates are undoubtedly one of the most diverse protozoans that play a significant role in ecology. However, molecular examination, based on comparing the DNA sequences, has been done on a limited number of the species. Because most ciliates are uncultivable and their population sizes are often too small, it is usually difficult to obtain sufficient genomic DNA required for PCR based experiments. In the present study, we evaluated the effectiveness of four commercial DNA extraction procedures that extract high quality genomic DNA from a single ciliate cell. It was discovered that RED Extract-N-$Amp^{TM}$ PCR kit is the best method for removing PCR-inhibiting substances and minimizing DNA loss during purification. This method can also amplify more than 25 reactions of PCR. In addition, this technique was applied to single cells of 19 species belonged to 7 orders under 5 classes that isolated from mixed natural populations. Their small subunit ribosomal DNA (SSU rDNA) was successfully amplified. In summary, we developed a simple technique for the high-yield extraction of purified DNA from a single ciliate cell that may be more useful for rare ciliates, such as tiny and uncultivable marine microbes.

• Journal of Food Hygiene and Safety
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• v.34 no.6
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• pp.557-561
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• 2019

This study investigated the efficacy of DNA extraction methods for real-time PCR detection of foodborne pathogenic bacteria in livestock manure composts. Livestock manure composts were inoculated with Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, Bacillus cereus and incubated in enrichment broth. For DNA extraction, enriched samples were treated following boiling method, by chloroform, C₁₈ powder, and proteinase K. As a result, 4 species of bacteria were detected by real-time PCR when subjected to boiling for 30 min and treated with proteinase K. These results suggest that detection of foodborne pathogens by real-time PCR from livestock manure composts could be applicable using effective DNA extraction methodology such as the boiling method or proteinase K.