• Korean Journal of Food Science and Technology
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• v.31 no.1
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• pp.110-114
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• 1999

The present study was conducted to investigate the preparative methods of agarose for gel electrophoresis from agar. Naturally occuring agar consists of two main polysaccharides, the neutral polysaccharide agarose and the acid sulphated polysaccharide agaropectin. The sulphate and carboxyl functions of the agar are accumulated in the agaropectin. The hydrophilic, non-ionogenic, rigid and transparent gel matrix of the agarose was found to be suitable for gel electrophoresis gel filtration and affinity chromatography. Agar was purified by chitosan treatment, cetylpyridinium chloride (CPC) treatment, and polyethylene glycol (PEG) treatment. Yields of agarose purified from agar with chitosan, CPC and PEG were 56.7%, 55.6% and 62.3%. It was proper to treat with chitosan in preparative methods of agarose for gel electrophoresis from agar.

• Korean Journal of Ichthyology
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• v.3 no.1
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• pp.58-62
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• 1991

The genomic DNA from liver tissue of rainbow trout(Oncorhynchus mykiss) was analyzed by agarose gel electrophoresis. The molecular weight of Oncorhynchus mykiss genomic DNA was about 21.3 kilobase pairs.

• The Korean Journal of Zoology
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• v.29 no.4
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• pp.283-293
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• 1986

For the preliminary step to make and characterize the monoclonal antibodies of human alpha-fetoprotein (AFP) was purified from 534g of human fetal tissues through the procedures of tissue extraction, DEAE-cellulose, concanavalin A-Sepharose, Cibacron Blue F3GA-agarose and immunoadsorbent affinity chromatography. The isolated AFP preparation showed a single band on polyacrylamide gel electrophoresis and a single precipitin are against rabbit anti-human cord serum and anti-human AFP on immunoelectrophoresis. Our AFP also displayed a single band on SDS-polyacrylamide gel electrophoresis. The recovery of AFP was 8.76mg total.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.6
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• pp.635-643
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• 2011

Agar was prepared from Gelidium amansii collected from Jeju Island, South Korea. This agar preparation has high gel strength and low sulfate content compared with G. amansii agar from Morocco. Accordingly, agarose was made from the Jeju agar through the consecutive refining processes of dimethyl sulfoxide (DMSO) extraction and ethylene diamine tetra acetic acid (EDTA) washing. The physicochemical properties of the resulting agarose were compared with those from agarose prepared using only DMSO extraction. Consecutive DMSO extraction and EDTA washing more strongly affected the physicochemical properties of the agarose (purified agarose) compared with the use of DMSO extraction alone. These properties were similar to those of commercial agarose used for electrophoresis. In DNA electrophoresis, the separation and movement speed of the purified agarose were similar to those of the commercial agarose. In a $^{13}C$ NMR analysis, the purified agarose exhibited the same carbon peak as the commercial agarose. When observed under scanning electron microscopy, the agar had an even and smooth surface without irregularities or pores, and the purified agarose had a wide surface area with a large number of pores; the commercial agarose had an irregular surface that would allow the solvent to easily permeate. These results illustrate that the physicochemical properties of agarose prepared from DMSO extraction and EDTA washing were more effective than those observed after DMSO extraction alone; thus, these processes used in succession will be useful in agarose industries.

• Environmental Mutagens and Carcinogens
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• v.17 no.2
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• pp.71-77
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• 1997

The single cell gel electrophoressis(SCGE) assay, also known as the comet assay, is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakage in mammalian cells. The SCGE or comet assay is a promising test for the detection of DNA damage and repair in individnal cells. It has widespread potential applications in DNA damage and repair studies, genotoxicity testing and biomonitoring. In this microgel electrophoresis technique, cells are embedded in agarose gel on microscope slides, iysed and electrophoresed under alkaline conditions. Cells with increased DNA damage display increased migration of DNA from the nucleus towards the anode. The length of DNA migration indicates the amount of DNA breakage in the cell. The comet assay is also capable of identifying apoptotic cells which contain highly fragmented DNA. Here we review the development of the SCGE assay, existing protocols for the detection and analysis of comets, the relevant underlying principles determining the behaviour of DNA and the potential applications of the technique.

• BMB Reports
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• v.38 no.6
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• pp.763-765
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• 2005

A new simplified procedure for identifying human plasmin was developed using a DTT copolymerized agarose stacking gel (ASG) system. Agarose (1%) was used for the stacking gel because DTT inhibits the polymerization of acrylamide. Human plasmin showed the lowest activity at pH 9.0. There was a similar catalytically active pattern observed under acidic conditions (pH 3.0) to that observed under alkaline conditions (pH 10.0 or 11.0). Using the ASG system, the primary structure of the heavy chain could be established at pH 3.0. This protein was found to consist of three fragments, 45 kDa, 23 kDa, and 13 kDa. These results showed that the heavy chain has a similar structure to the autolysed plasmin (Wu et al., 1987b) but there is a different start amino acid sequence of the N-termini.

• Journal of Radiation Protection and Research
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• v.31 no.2
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• pp.65-68
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• 2006

In this study, the extent of plasmid DNA damage was observed according to concentration of BSH(Boron Sulfhydryl Hydride) and irradiation doses of neutron and ${\gamma}$-ray. The plasmid used was both pBR 322 (2870 bp) and ${\Phi}X174$ RF(5386 bp) DNA. Plasmid DNA damage by irradiation in the presence of BSH was analyzed by agarose gel electrophoresis. In the neutron experiment, DNA damage of both plasmid DNAs was increased according to increasing the concentration of BSH and neutron doses. But in the ${\gamma}$-ray experiment, there appeared no dose dependency as compared to the neutron experiment. The extent of the plasmid DNA damage in the presence of BSH was somewhat different according to irradiation by neutron or ${\gamma}$-ray.

• Mycobiology
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• v.39 no.1
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• pp.45-51
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• 2011

This work was conducted to investigate dietary supplementation of oyster mushroom fruiting bodies on biochemical and histological changes in hyper and normocholesterolemic rats. Six-week old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. Feeding a diet containing a 5% powder of Pleurotus ostreatus fruiting bodies to hypercholesterolemic rats reduced plasma total cholesterol, triglyceride, low-density lipoprotein (LDL), total lipid, phospholipids, and LDL/high-density lipoprotein ratio by 30.18, 52.75, 59.62, 34.15, 23.89, and 50%, respectively. Feeding oyster mushrooms also significantly reduced body weight in hypercholesterolemic rats. However, it had no adverse effects on plasma albumin, total bilirubin, direct bilirubin, creatinin, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, or enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that P. ostreatus significantly reduced plasma ${\beta}$ and pre-${\beta}$-lipoprotein but increased ${\alpha}$-lipoprotein. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red O staining revealed normal findings for mushroom-fed hypercholesterolemic rats. These results suggest that a 5% P. ostreatus diet supplement provided health benefits by acting on the atherogenic lipid profile in hypercholesterolemic rats.

• Toxicological Research
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• v.25 no.3
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• pp.113-118
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• 2009

Cell death induced by menadione (vitamin K-3,2-methyl-1,4-naphthoquinone) has been investigated in human promyelocytic leukemia HL-60 cells. Menadione was found to induce both apoptosis and necrosis in HL-60 cells. Low concentration ($1{\sim}$50 ${\mu}$M) of menadione induced apoptotic cell death, which was demonstrated by typical DNA ladder patterns on agarose gel electrophoresis and flow cytometry analysis. In contrast, a high concentration of menadione (100 ${\mu}$M) induced necrotic cell death, which was demonstrated by DNA smear pattern in agarose gel electrophoresis. Necrotic cell death was accompanied with a great reduction of cell viability. Menadione activated caspase-3, as evidenced by both increased protease activity and proteolytic cleavage of 116 kDa poly(ADP-ribose) polymerase (PARP) into 85 kDa cleavage product. Caspase-3 activity was maximum at 50 ${\mu}$M of menadione, and very low at 100 ${\mu}$M of menadione. Taken together, our results showed that menadione induced mixed types of cell death, apoptosis at low concentrations and necrosis at high concentrations in HL-60 cells.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.359-363
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• 2004

I investigated an effective way to generate a metagenomic library from DNA prepared from environental samples. The sizes of DNA extracted from environmental samples were usually in the range of 10 to 100 kbp as estimated from $0.4\%$ agarose gel electrophoresis. Because of this small size, a fosmid, rather than BAC, was chosen as a vector. It was found that, for the successful generation of metagenomic library, the selection of DNA with the sized of about 40 kbp was critical and, therefore, a simple agarose gel electrophoresis system was developed to select this size of DNA. By the procedure described in this report, I obtained metagenomic libraries containing 25,000 fosmid clones, which corresponded to 1,000 Mb of metagenomic DNA.