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Bio-Rad tbe buffer
Electrophoresis mobility shift assay (EMSA) and DNA footprinting assays indicate HlyU binding to a region within the exsA promoter. (A) Six percent <t>TBE-polyacrylamide</t> gels were loaded with reaction mixtures containing exsA promoter DNA and increasing amounts of purified HlyU-His or bovine serum albumin (BSA). An exsA promoter DNA-only control was included as indicated. The white box indicates slow-migrating, concentrated DNA species. (B) A 6% TBE-polyacrylamide gel was loaded with reaction mixtures containing nleH1 DNA and increasing amounts of purified HlyU-His. Following electrophoresis, gels were stained with <t>SYBR</t> green to specifically stain DNA. (C) A DNase I footprinting assay using purified HlyU-His was used to identify a DNA binding region for HlyU-His within the exsA promoter. The approximate base pair numbers of the HlyU-His footprint region are indicated (259 to 315 bp) and are based on capillary electrophoresis internal size standards. The DNA sequence associated with the identified HlyU protected region is displayed below the chromatogram. A 7-bp inverted repeat (labeled in blue) and separated by 14 bp is centrally located within the HlyU-His protected region.
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1) Product Images from "The Transcriptional Regulator HlyU Positively Regulates Expression of exsA, Leading to Type III Secretion System 1 Activation in Vibrio parahaemolyticus"

Article Title: The Transcriptional Regulator HlyU Positively Regulates Expression of exsA, Leading to Type III Secretion System 1 Activation in Vibrio parahaemolyticus

Journal: Journal of Bacteriology

doi: 10.1128/JB.00653-17

Electrophoresis mobility shift assay (EMSA) and DNA footprinting assays indicate HlyU binding to a region within the exsA promoter. (A) Six percent TBE-polyacrylamide gels were loaded with reaction mixtures containing exsA promoter DNA and increasing amounts of purified HlyU-His or bovine serum albumin (BSA). An exsA promoter DNA-only control was included as indicated. The white box indicates slow-migrating, concentrated DNA species. (B) A 6% TBE-polyacrylamide gel was loaded with reaction mixtures containing nleH1 DNA and increasing amounts of purified HlyU-His. Following electrophoresis, gels were stained with SYBR green to specifically stain DNA. (C) A DNase I footprinting assay using purified HlyU-His was used to identify a DNA binding region for HlyU-His within the exsA promoter. The approximate base pair numbers of the HlyU-His footprint region are indicated (259 to 315 bp) and are based on capillary electrophoresis internal size standards. The DNA sequence associated with the identified HlyU protected region is displayed below the chromatogram. A 7-bp inverted repeat (labeled in blue) and separated by 14 bp is centrally located within the HlyU-His protected region.
Figure Legend Snippet: Electrophoresis mobility shift assay (EMSA) and DNA footprinting assays indicate HlyU binding to a region within the exsA promoter. (A) Six percent TBE-polyacrylamide gels were loaded with reaction mixtures containing exsA promoter DNA and increasing amounts of purified HlyU-His or bovine serum albumin (BSA). An exsA promoter DNA-only control was included as indicated. The white box indicates slow-migrating, concentrated DNA species. (B) A 6% TBE-polyacrylamide gel was loaded with reaction mixtures containing nleH1 DNA and increasing amounts of purified HlyU-His. Following electrophoresis, gels were stained with SYBR green to specifically stain DNA. (C) A DNase I footprinting assay using purified HlyU-His was used to identify a DNA binding region for HlyU-His within the exsA promoter. The approximate base pair numbers of the HlyU-His footprint region are indicated (259 to 315 bp) and are based on capillary electrophoresis internal size standards. The DNA sequence associated with the identified HlyU protected region is displayed below the chromatogram. A 7-bp inverted repeat (labeled in blue) and separated by 14 bp is centrally located within the HlyU-His protected region.

Techniques Used: Electrophoresis, Mobility Shift, DNA Footprinting, Binding Assay, Purification, Staining, SYBR Green Assay, Footprinting, Sequencing, Labeling

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Polyacrylamide Gel Electrophoresis:

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Electrophoresis:

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Agarose Gel Electrophoresis:

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Article Title: Quantitative analysis of TALE-DNA interactions suggests polarity effects
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Article Title: Reconstitution of Drosophila and human chromatins by wheat germ cell-free co-expression system
Article Snippet: Supercoiling assayFifty-five point zero microliter of the total chromatin assembly reaction mixture was purified by phenol-chloroform-IAA (25:24:1, v/v) extraction (pH 8.0) followed by ethanol precipitation, and the purified DNA was resuspended in HD buffer (25 mM HEPES, 1 mM DTT, pH 7.6) containing a trace amount of Ribonuclease a (Macherey-Nagel GmbH & Co.). .. Supercoiled plasmid DNAs were separated by 0.8% TBE agarose gel in 0.5x TBE buffer, visualized with ethidium bromide (Nippon Gene), and analyzed by gel documentation system (Bio-Rad Laboratories). .. Micrococcal nuclease assay MNase assay was performed by the addition of Micrococcal Nuclease (Bio-Rad Laboratories) and MNase buffer (20 mM Tris-HCl (pH 8.0), 5 mM NaCl, 2.5 mM CaCl2 ) to 28.0 μl of total reaction mixture followed by 5, 10, 20, 30, 40 min incubation at 25 °C.

Size-exclusion Chromatography:

Article Title: A Sequence-Ready BAC Contig of the GABAA Receptor Gene Cluster Gabrg1–Gabra2–Gabrb1 on Mouse Chromosome 5
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BAC Assay:

Article Title: A Sequence-Ready BAC Contig of the GABAA Receptor Gene Cluster Gabrg1–Gabra2–Gabrb1 on Mouse Chromosome 5
Article Snippet: Miniprep DNA (5 μl) was digested immediately in a total volume of 20 μl with 5 units of Not I enzyme (New England Biolabs, Inc., Beverly, MA) for 2 hr at 37°C. .. Samples were loaded on a 1% agarose gel in 0.5% Tris-borate–EDTA (TBE) and subjected to PFGE (Bio-Rad CHEF DR II) for 16 hr at 6 V/cm, 15°C with a switching interval from 5 sec to 15 sec. BAC insert sizes were assigned from ethidium bromide-stained gels using AlphEase software and an Alpha-Imager 2000 gel-documentation system (Alpha Innotech, San Leandro, CA). .. Eco RI digests of freshly prepared miniprep DNA were also used to fingerprint clones according to the methods of .

Software:

Article Title: A Sequence-Ready BAC Contig of the GABAA Receptor Gene Cluster Gabrg1–Gabra2–Gabrb1 on Mouse Chromosome 5
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Article Title: Morphological and molecular datasets for Kaempferia species
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Binding Assay:

Article Title: Quantitative analysis of TALE-DNA interactions suggests polarity effects
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Nucleic Acid Electrophoresis:

Article Title: Quantitative analysis of TALE-DNA interactions suggests polarity effects
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Plasmid Preparation:

Article Title: Reconstitution of Drosophila and human chromatins by wheat germ cell-free co-expression system
Article Snippet: Supercoiling assayFifty-five point zero microliter of the total chromatin assembly reaction mixture was purified by phenol-chloroform-IAA (25:24:1, v/v) extraction (pH 8.0) followed by ethanol precipitation, and the purified DNA was resuspended in HD buffer (25 mM HEPES, 1 mM DTT, pH 7.6) containing a trace amount of Ribonuclease a (Macherey-Nagel GmbH & Co.). .. Supercoiled plasmid DNAs were separated by 0.8% TBE agarose gel in 0.5x TBE buffer, visualized with ethidium bromide (Nippon Gene), and analyzed by gel documentation system (Bio-Rad Laboratories). .. Micrococcal nuclease assay MNase assay was performed by the addition of Micrococcal Nuclease (Bio-Rad Laboratories) and MNase buffer (20 mM Tris-HCl (pH 8.0), 5 mM NaCl, 2.5 mM CaCl2 ) to 28.0 μl of total reaction mixture followed by 5, 10, 20, 30, 40 min incubation at 25 °C.

Staining:

Article Title: The Transcriptional Regulator HlyU Positively Regulates Expression of exsA, Leading to Type III Secretion System 1 Activation in Vibrio parahaemolyticus
Article Snippet: .. The gel was run for 4 h (100 V, 4°C) and then stained with SYBR green fluorescent DNA dye (Invitrogen) at a 1× concentration in TBE buffer and imaged using a VersaDoc MP5000 system (Bio-Rad). .. Protein staining and processing of TBE gels was performed using SYPRO Ruby Red (Bio-Rad) as previously described ( ) and then imaged using a VersaDoc MP5000 system (Bio-Rad).

Article Title: Morphological and molecular datasets for Kaempferia species
Article Snippet: Each PCR reaction was in a total volume of 25 µL, containing 15 ng of template DNA, 12.5 µL of 2x Type-it PCR Master Mix (Red-Taq), 1 µL each of 10 µM ITS4 and ITS5, and 8.5 µL of RNase-free water (Qiagen®). .. Amplification products were separated via electrophoresis on 1.5% (w/v) agarose gels with 1x TBE buffer at 70 V for 75 min, stained with GelRedTM Nucleic Acid Stain and visualized under UV light using Bio-Rad Molecular Imager GelDocTM XR+ with Image LabTM Software (Bio-Rad Laboratories, Inc.,USA) ( ). .. GeneRuler 100 bp ladder (Fermentas) was used as DNA molecular weight markers.

SYBR Green Assay:

Article Title: The Transcriptional Regulator HlyU Positively Regulates Expression of exsA, Leading to Type III Secretion System 1 Activation in Vibrio parahaemolyticus
Article Snippet: .. The gel was run for 4 h (100 V, 4°C) and then stained with SYBR green fluorescent DNA dye (Invitrogen) at a 1× concentration in TBE buffer and imaged using a VersaDoc MP5000 system (Bio-Rad). .. Protein staining and processing of TBE gels was performed using SYPRO Ruby Red (Bio-Rad) as previously described ( ) and then imaged using a VersaDoc MP5000 system (Bio-Rad).

Concentration Assay:

Article Title: The Transcriptional Regulator HlyU Positively Regulates Expression of exsA, Leading to Type III Secretion System 1 Activation in Vibrio parahaemolyticus
Article Snippet: .. The gel was run for 4 h (100 V, 4°C) and then stained with SYBR green fluorescent DNA dye (Invitrogen) at a 1× concentration in TBE buffer and imaged using a VersaDoc MP5000 system (Bio-Rad). .. Protein staining and processing of TBE gels was performed using SYPRO Ruby Red (Bio-Rad) as previously described ( ) and then imaged using a VersaDoc MP5000 system (Bio-Rad).

Amplification:

Article Title: Morphological and molecular datasets for Kaempferia species
Article Snippet: Each PCR reaction was in a total volume of 25 µL, containing 15 ng of template DNA, 12.5 µL of 2x Type-it PCR Master Mix (Red-Taq), 1 µL each of 10 µM ITS4 and ITS5, and 8.5 µL of RNase-free water (Qiagen®). .. Amplification products were separated via electrophoresis on 1.5% (w/v) agarose gels with 1x TBE buffer at 70 V for 75 min, stained with GelRedTM Nucleic Acid Stain and visualized under UV light using Bio-Rad Molecular Imager GelDocTM XR+ with Image LabTM Software (Bio-Rad Laboratories, Inc.,USA) ( ). .. GeneRuler 100 bp ladder (Fermentas) was used as DNA molecular weight markers.

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    Optimization and confirmation of human chromatin assembly reaction. a DNA supercoiling assay to optimize the ratio of HsH2A/HsH2B and HsH3.1/HsH4 mRNAs in the chromatin reconstitution reaction for 4 h. b Supercoiling assay of assembled human chromatin. Incubation time indicated with 0, 1, 4, 6, and 8 h, where 0 indicated the reconstitution reaction in the absence of mRNAs encoding histones. Supercoils were analyzed by 0.8% agarose <t>TBE</t> gel. Relaxed, linear, and <t>supercoiled</t> DNA are indicated as RC, L, and SC, respectively. c Partial MNase digestion of assembled human chromatin, then samples were run on 2.0% agarose gel. Reconstituted chromatin was digested by MNase for the indicated time (0, 5, 10, 20, 30, and 40 min), and bands corresponding to digestion fragment of the mono-, di-, tri- and tetranucleosome were indicated. Mr: DNA molecular weight marker
    Tbe Buffer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The complex role of iron in CN+HP toxicity A. CN inhibits the standard Fenton reaction in vitro via formation of Fe(CN) 6 . The reaction, which is started by addition of FeSO 4 to the indicated concentration to solution containing 2 mM HP, is followed spectrophotometrically (absorbance at 440 nm), by disappearance of a colored substance, p -nitrosodimethylaniline (p-NDA). When added, CN is at 3 mM. B. Pre-treatment of the cultures with iron chelators, such as 20 mM deferoxamine or 2 mM dipyridyl, for five minutes before the treatment blocks CN+HP killing. C. Pre-treatment with the iron chelators like in “B” similarly blocks CN+HP-induced chromosome fragmentation. D. CN inhibits Fenton-like in vitro reactions with other transition metals. The reactions contained 50 μM <t>EDTA</t> and 250 μM of the indicated metal and were ran like in “A”. The values are means of two independent repetitions. E. CN stimulates in vitro Fenton’s reaction with free iron in water or a <t>Tris</t> buffer and plasmid relaxation as a readout. SCD, supercoiled dimer; SCM, supercoiled monomer; RCD, relaxed circular dimer; RCM, relaxed circular monomer. Concentrations were: 250 μM FeSO 4 , 2 mM HP, 3 mM CN, 40 mM Tris-HCl pH 8.0. F. Quantification of several gels like in “E”. The values are (RCM/RCM+SCM)×100. G. Kinetics of killing by the two treatments of the fur mutant. H. Kinetics of killing by the two treatments of the sodAB double mutant. I. Our original idea about free cytoplasmic iron (top) and its current evolution (bottom). Small green circles, DNA-accessible Fe(II); orange and brown hexagons, Fe(III)-depots.
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    Optimization and confirmation of human chromatin assembly reaction. a DNA supercoiling assay to optimize the ratio of HsH2A/HsH2B and HsH3.1/HsH4 mRNAs in the chromatin reconstitution reaction for 4 h. b Supercoiling assay of assembled human chromatin. Incubation time indicated with 0, 1, 4, 6, and 8 h, where 0 indicated the reconstitution reaction in the absence of mRNAs encoding histones. Supercoils were analyzed by 0.8% agarose TBE gel. Relaxed, linear, and supercoiled DNA are indicated as RC, L, and SC, respectively. c Partial MNase digestion of assembled human chromatin, then samples were run on 2.0% agarose gel. Reconstituted chromatin was digested by MNase for the indicated time (0, 5, 10, 20, 30, and 40 min), and bands corresponding to digestion fragment of the mono-, di-, tri- and tetranucleosome were indicated. Mr: DNA molecular weight marker

    Journal: BMC Biotechnology

    Article Title: Reconstitution of Drosophila and human chromatins by wheat germ cell-free co-expression system

    doi: 10.1186/s12896-020-00655-6

    Figure Lengend Snippet: Optimization and confirmation of human chromatin assembly reaction. a DNA supercoiling assay to optimize the ratio of HsH2A/HsH2B and HsH3.1/HsH4 mRNAs in the chromatin reconstitution reaction for 4 h. b Supercoiling assay of assembled human chromatin. Incubation time indicated with 0, 1, 4, 6, and 8 h, where 0 indicated the reconstitution reaction in the absence of mRNAs encoding histones. Supercoils were analyzed by 0.8% agarose TBE gel. Relaxed, linear, and supercoiled DNA are indicated as RC, L, and SC, respectively. c Partial MNase digestion of assembled human chromatin, then samples were run on 2.0% agarose gel. Reconstituted chromatin was digested by MNase for the indicated time (0, 5, 10, 20, 30, and 40 min), and bands corresponding to digestion fragment of the mono-, di-, tri- and tetranucleosome were indicated. Mr: DNA molecular weight marker

    Article Snippet: Supercoiled plasmid DNAs were separated by 0.8% TBE agarose gel in 0.5x TBE buffer, visualized with ethidium bromide (Nippon Gene), and analyzed by gel documentation system (Bio-Rad Laboratories).

    Techniques: Incubation, Agarose Gel Electrophoresis, Molecular Weight, Marker

    DNA supercoiling and MNase assays of Drosophila chromatin assembly reactions. a Determination of the appropriate ratio of DmH2A/DmH2B and DmH3/DmH4 mRNAs in the Drosophila chromatin reconstitution reaction. The incubation time was 4 h. Supercoiling assay was performed, samples were analyzed by 0.8% agarose TBE gel. Relaxed, linear, and supercoiled plasmid DNAs are indicated as RC, L, and SC, respectively. b Supercoiling assay of assembled Drosophila chromatin under the indicated reaction time 0, 1, 4, and 6 h, where 0 indicated the reconstitution reaction in the absence of mRNAs encoding histones. Supercoiling assay was performed, samples were analyzed by 0.8% agarose TBE gel. c Partial MNase digestion was performed on assembled Drosophila chromatin for the indicated time (0, 5, 10, 20, 30, and 40 min), then samples were run on 2.0% agarose gel. Bands corresponding to digestion fragment of the mono-, di-, and trinucleosomes were detected and indicated. Agarose gels were visualized by ethidium bromide and UV light. Mr: DNA molecular weight marker

    Journal: BMC Biotechnology

    Article Title: Reconstitution of Drosophila and human chromatins by wheat germ cell-free co-expression system

    doi: 10.1186/s12896-020-00655-6

    Figure Lengend Snippet: DNA supercoiling and MNase assays of Drosophila chromatin assembly reactions. a Determination of the appropriate ratio of DmH2A/DmH2B and DmH3/DmH4 mRNAs in the Drosophila chromatin reconstitution reaction. The incubation time was 4 h. Supercoiling assay was performed, samples were analyzed by 0.8% agarose TBE gel. Relaxed, linear, and supercoiled plasmid DNAs are indicated as RC, L, and SC, respectively. b Supercoiling assay of assembled Drosophila chromatin under the indicated reaction time 0, 1, 4, and 6 h, where 0 indicated the reconstitution reaction in the absence of mRNAs encoding histones. Supercoiling assay was performed, samples were analyzed by 0.8% agarose TBE gel. c Partial MNase digestion was performed on assembled Drosophila chromatin for the indicated time (0, 5, 10, 20, 30, and 40 min), then samples were run on 2.0% agarose gel. Bands corresponding to digestion fragment of the mono-, di-, and trinucleosomes were detected and indicated. Agarose gels were visualized by ethidium bromide and UV light. Mr: DNA molecular weight marker

    Article Snippet: Supercoiled plasmid DNAs were separated by 0.8% TBE agarose gel in 0.5x TBE buffer, visualized with ethidium bromide (Nippon Gene), and analyzed by gel documentation system (Bio-Rad Laboratories).

    Techniques: Incubation, Plasmid Preparation, Agarose Gel Electrophoresis, Molecular Weight, Marker

    The complex role of iron in CN+HP toxicity A. CN inhibits the standard Fenton reaction in vitro via formation of Fe(CN) 6 . The reaction, which is started by addition of FeSO 4 to the indicated concentration to solution containing 2 mM HP, is followed spectrophotometrically (absorbance at 440 nm), by disappearance of a colored substance, p -nitrosodimethylaniline (p-NDA). When added, CN is at 3 mM. B. Pre-treatment of the cultures with iron chelators, such as 20 mM deferoxamine or 2 mM dipyridyl, for five minutes before the treatment blocks CN+HP killing. C. Pre-treatment with the iron chelators like in “B” similarly blocks CN+HP-induced chromosome fragmentation. D. CN inhibits Fenton-like in vitro reactions with other transition metals. The reactions contained 50 μM EDTA and 250 μM of the indicated metal and were ran like in “A”. The values are means of two independent repetitions. E. CN stimulates in vitro Fenton’s reaction with free iron in water or a Tris buffer and plasmid relaxation as a readout. SCD, supercoiled dimer; SCM, supercoiled monomer; RCD, relaxed circular dimer; RCM, relaxed circular monomer. Concentrations were: 250 μM FeSO 4 , 2 mM HP, 3 mM CN, 40 mM Tris-HCl pH 8.0. F. Quantification of several gels like in “E”. The values are (RCM/RCM+SCM)×100. G. Kinetics of killing by the two treatments of the fur mutant. H. Kinetics of killing by the two treatments of the sodAB double mutant. I. Our original idea about free cytoplasmic iron (top) and its current evolution (bottom). Small green circles, DNA-accessible Fe(II); orange and brown hexagons, Fe(III)-depots.

    Journal: Molecular microbiology

    Article Title: Cyanide enhances hydrogen peroxide toxicity by recruiting endogenous iron to trigger catastrophic chromosomal fragmentation

    doi: 10.1111/mmi.12938

    Figure Lengend Snippet: The complex role of iron in CN+HP toxicity A. CN inhibits the standard Fenton reaction in vitro via formation of Fe(CN) 6 . The reaction, which is started by addition of FeSO 4 to the indicated concentration to solution containing 2 mM HP, is followed spectrophotometrically (absorbance at 440 nm), by disappearance of a colored substance, p -nitrosodimethylaniline (p-NDA). When added, CN is at 3 mM. B. Pre-treatment of the cultures with iron chelators, such as 20 mM deferoxamine or 2 mM dipyridyl, for five minutes before the treatment blocks CN+HP killing. C. Pre-treatment with the iron chelators like in “B” similarly blocks CN+HP-induced chromosome fragmentation. D. CN inhibits Fenton-like in vitro reactions with other transition metals. The reactions contained 50 μM EDTA and 250 μM of the indicated metal and were ran like in “A”. The values are means of two independent repetitions. E. CN stimulates in vitro Fenton’s reaction with free iron in water or a Tris buffer and plasmid relaxation as a readout. SCD, supercoiled dimer; SCM, supercoiled monomer; RCD, relaxed circular dimer; RCM, relaxed circular monomer. Concentrations were: 250 μM FeSO 4 , 2 mM HP, 3 mM CN, 40 mM Tris-HCl pH 8.0. F. Quantification of several gels like in “E”. The values are (RCM/RCM+SCM)×100. G. Kinetics of killing by the two treatments of the fur mutant. H. Kinetics of killing by the two treatments of the sodAB double mutant. I. Our original idea about free cytoplasmic iron (top) and its current evolution (bottom). Small green circles, DNA-accessible Fe(II); orange and brown hexagons, Fe(III)-depots.

    Article Snippet: Half plugs were loaded into a 1.0% agarose gel in 0.5× Tris-borate-EDTA buffer and run at 6.0 V/cm with the initial and the final switch times of 60 and 120 s, respectively, at 12°C in a Bio-Rad CHEF-DR II PFGE system for 20-22 hours.

    Techniques: In Vitro, Concentration Assay, Plasmid Preparation, Mutagenesis

    PFGE patterns of recently isolated O3:K6 strains of V. parahaemolyticus . Conditions for PFGE were 1% agarose gel, 0.5× Tris-borate-EDTA buffer, 190 V, pulse time 3 to 80 s, for 22.4 h. Lane 1, isolate 1020 (from Philippines; pattern I5); lane 2, isolate 1021 (from Singapore; pattern I6); lane 3, isolate 1084 (from Taiwan; pattern I7); lane 4, isolate 1104 (from Taiwan; pattern I8); lane 5, isolate 1123 (from Taiwan; pattern I1); lane 6, isolate 1125 (from Taiwan; pattern I1); lane 7, isolate 1139 (from Taiwan; pattern I4); lane 8, isolate 1154 (from Taiwan; pattern I2); lane 9, isolate 97-804 (from Korea; pattern I3); lane M, lambda ladder PFGE marker.

    Journal: Applied and Environmental Microbiology

    Article Title: Characteristics of Vibrio parahaemolyticus O3:K6 from Asia

    doi:

    Figure Lengend Snippet: PFGE patterns of recently isolated O3:K6 strains of V. parahaemolyticus . Conditions for PFGE were 1% agarose gel, 0.5× Tris-borate-EDTA buffer, 190 V, pulse time 3 to 80 s, for 22.4 h. Lane 1, isolate 1020 (from Philippines; pattern I5); lane 2, isolate 1021 (from Singapore; pattern I6); lane 3, isolate 1084 (from Taiwan; pattern I7); lane 4, isolate 1104 (from Taiwan; pattern I8); lane 5, isolate 1123 (from Taiwan; pattern I1); lane 6, isolate 1125 (from Taiwan; pattern I1); lane 7, isolate 1139 (from Taiwan; pattern I4); lane 8, isolate 1154 (from Taiwan; pattern I2); lane 9, isolate 97-804 (from Korea; pattern I3); lane M, lambda ladder PFGE marker.

    Article Snippet: It was then incubated at 4°C for 16 h and, finally, digestion was performed at 37°C for another 48 h. High-molecular-weight restriction fragments were resolved in 1% agarose gel in 0.5% Tris-borate-EDTA buffer by using a CHEF apparatus (CHEF-DR II; Bio-Rad Laboratories, Richmond, Calif.).

    Techniques: Isolation, Agarose Gel Electrophoresis, Marker