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  • 95
    Integrated DNA Technologies rhamp snp genotyping system
    Allelic discrimination plots obtained for SNP103 using TaqMan, KASP and <t>rhAmp</t> <t>genotyping</t> assays on 96 samples. Red and blue dots represent the homozygous genotypes, the green circles represent heterozygous genotypes and the squares on the bottom left of the plot are no-template control
    Rhamp Snp Genotyping System, supplied by Integrated DNA Technologies, used in various techniques. Bioz Stars score: 95/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore rsa
    In vivo biodistribution of <t>ABD-fused</t> (Z HER2:342 ) 2 in a preclinical rat model. A , ELISA measurements showing pH-dependent binding of srFcRn to <t>RSA</t> in complex with ABD-(Z HER2:342 ) 2 . The numbers given represent the mean of triplicates. The blood, skin, and muscle biodistribution of 111 In-labeled-RSA ( B ) and 177 Lu-labeled-ABD-(Z HER2:342 ) 2 ( C ) in Sprague-Dawley rats (three rats/group and time point). Organ uptake is expressed as percent of injected radioactivity/g (% IA/g ), and error bars and corrected for interstitial volume as described under “Experimental Procedures.”
    Rsa, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 150 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    TA Instruments g2 rsa
    In vivo biodistribution of <t>ABD-fused</t> (Z HER2:342 ) 2 in a preclinical rat model. A , ELISA measurements showing pH-dependent binding of srFcRn to <t>RSA</t> in complex with ABD-(Z HER2:342 ) 2 . The numbers given represent the mean of triplicates. The blood, skin, and muscle biodistribution of 111 In-labeled-RSA ( B ) and 177 Lu-labeled-ABD-(Z HER2:342 ) 2 ( C ) in Sprague-Dawley rats (three rats/group and time point). Organ uptake is expressed as percent of injected radioactivity/g (% IA/g ), and error bars and corrected for interstitial volume as described under “Experimental Procedures.”
    G2 Rsa, supplied by TA Instruments, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Boehringer Mannheim rsa i
    In vivo biodistribution of <t>ABD-fused</t> (Z HER2:342 ) 2 in a preclinical rat model. A , ELISA measurements showing pH-dependent binding of srFcRn to <t>RSA</t> in complex with ABD-(Z HER2:342 ) 2 . The numbers given represent the mean of triplicates. The blood, skin, and muscle biodistribution of 111 In-labeled-RSA ( B ) and 177 Lu-labeled-ABD-(Z HER2:342 ) 2 ( C ) in Sprague-Dawley rats (three rats/group and time point). Organ uptake is expressed as percent of injected radioactivity/g (% IA/g ), and error bars and corrected for interstitial volume as described under “Experimental Procedures.”
    Rsa I, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    GE Healthcare rsa i
    Neighbour-joining similarity tree for 16S rRNA gene sequences of most mycobacterial species. Legend: N. asteroides ATCC 49872 (Genbank Z82229) was used as the outgroup. Table 1 lists the GenBank accession numbers of the sequences used to construct this tree. ARDRA patterns for Hha I, <t>Mbo</t> I, <t>Rsa</t> I and Bst UI are listed after the species name. a: GenBank AF028712. Erroneously listed in GenBank as M. peregrinum (see also legend of Table 1 ). b: M. gastri clusters below 100% with M. kansasii, although it is generally agreed that the 16S rRNA gene sequences for M. kansasii and M. gastri are identical. This can be explained by the fact that the only available GenBank M. gastri sequence (X52919) contained several ambiguities. c. M. lentiflavum, initially not included in the manuscript is not presented in this tree. It clusters close to the branch including M. heidelbergense, M. simiae, M. triplex and M. genavense.
    Rsa I, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Promega rsa i
    Representative diversity revealed by M46 PCR-RFLP in M. catarrhalis . (A) Amplimer B; (B) amplimer C. Samples were treated with Hae <t>III</t> (lanes 1 to 7) or with <t>Rsa</t> I (lanes 8 to 14) or are undigested amplimers (lanes 15 to 21). Each lane contained 10 μl of PCR product. Lanes 1, 8, and 15, sample ATCC 25238; lanes 2, 9, and 16, sample TN16a; lanes 3, 10, and 17, sample BL2; lanes 4, 11, and 18, sample NY1a; lanes 5, 12, and 19, sample TN17a; lanes 6, 13, and 20, sample TN7a; lanes 7, 14, and 21, sample TN13a. Lanes M are 100-to-1,500-bp ladders, with double-bright bands at 600 bp and additional fragments of 2,000 bp.
    Rsa I, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 567 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rsa i  (Roche)
    93
    Roche rsa i
    Relationship between mean telomere length, measured by Southern blots of the TRFs, versus the ratio of T (telomere amount)/Dx (DNA amount), measured by dot blots, and versus the ratio of T (telomere product)/S(single gene product), measured by qPCR. ( a and b ) The TRF products generated by <t>Hinf</t> I/ <t>Rsa</t> I and by Hph I/ Mnl I versus the T/Dx. ( c and d ) The TRF product generated by Hinf I/ Rsa I and by Hph I/ Mnl I versus T/S.
    Rsa I, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 249 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher rsa i
    A PCR-RFLP assay of the ITS2 region applied to pearls from Pinctada margaritifera , P. maxima and P. radiata . (A) PCR products of 575 bp ( P. margaritifera ), 571 bp ( P. maxima ) and 590–91 bp ( P. radiata ) obtained with ITS2 universal primers (5.8S-F and 28S-R) and (B) RFLP patterns of ITS2 amplicons (from A) obtained after digestion with <t>Rsa</t> I. MW: molecular weight size marker, 100-bp DNA ladder; lanes 1–3: P. maxima (PMX) pearls; lane 4: P. margaritifera (PMR) pearl; lanes 5–10: P. radiata (PR) pearls; lanes 11–16: P. margaritifera pearls; lane 17: PCR negative control; lanes 18 and 19: P. radiata and P. margaritifera positive controls. Note: The P. maxima positive control is shown in Figure 4 .
    Rsa I, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 482 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Difco rsa broth
    A PCR-RFLP assay of the ITS2 region applied to pearls from Pinctada margaritifera , P. maxima and P. radiata . (A) PCR products of 575 bp ( P. margaritifera ), 571 bp ( P. maxima ) and 590–91 bp ( P. radiata ) obtained with ITS2 universal primers (5.8S-F and 28S-R) and (B) RFLP patterns of ITS2 amplicons (from A) obtained after digestion with <t>Rsa</t> I. MW: molecular weight size marker, 100-bp DNA ladder; lanes 1–3: P. maxima (PMX) pearls; lane 4: P. margaritifera (PMR) pearl; lanes 5–10: P. radiata (PR) pearls; lanes 11–16: P. margaritifera pearls; lane 17: PCR negative control; lanes 18 and 19: P. radiata and P. margaritifera positive controls. Note: The P. maxima positive control is shown in Figure 4 .
    Rsa Broth, supplied by Difco, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Kaneka Corp rsa i
    ( a ) Western blot analysis of PC1 and PC2 protein expression in human islets cultured for 48 hours in the presence or absence of IL-1β (50 U/mL), IFN-γ (1,000 U/mL), or IL-1β plus IFN-γ. ( b ) Competitive <t>PCR</t> analysis of PC1 and PC2 mRNA expression in control and cytokine-cultured (IL-1β + IFN-γ, 48 hours) human islets. Lanes 1–4 contain the same amount of cDNA from the same human islet preparation and a decreasing amount of competitor DNA (0.5, 0.125, 0.03, and 0.0075 amol, respectively). Competitor DNA for PC2 was prepared from rat cDNA for PC1 from human cDNA. After amplification, PC1 products were directly run on 2% agarose-ethidium bromide gel, whereas PC2 products were digested with <t>Rsa</t> I before separation. Competitor rat PC2 appears as 2 bands (151 and 65 bp) after Rsa I restriction, but human PC2 remains intact. The figures are representative of 3 independent experiments.
    Rsa I, supplied by Kaneka Corp, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Toyobo rsa i
    Amplified products of the 16S rDNA of various Alex-CFPB (A) and digestion of the PCR products with <t>Rsa</t> I (B), Bst UI (C), <t>Mbo</t> I (D), and Hha I (E). Lane M, 100-bp ladder markers; lane 1, CFPB-A9; lane 2, CFPB-B1; lane 3, CFPB-C1; lane 4, CFPB-D1; lane 5, CFPB-E1; lane 6, CFPB-F1; lane 7, CFPB-G1; lane 8, CFPB-H1; lane 9, CFPB-I1; lane 10, CFPB-J1; lane 11, CFPB-K1; lane 12, CFPB-L1; lane 13, CFPB-M1; lane 14, CFPB-N1.
    Rsa I, supplied by Toyobo, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    TaKaRa rsa i
    Amplified products of the 16S rDNA of various Alex-CFPB (A) and digestion of the PCR products with <t>Rsa</t> I (B), Bst UI (C), <t>Mbo</t> I (D), and Hha I (E). Lane M, 100-bp ladder markers; lane 1, CFPB-A9; lane 2, CFPB-B1; lane 3, CFPB-C1; lane 4, CFPB-D1; lane 5, CFPB-E1; lane 6, CFPB-F1; lane 7, CFPB-G1; lane 8, CFPB-H1; lane 9, CFPB-I1; lane 10, CFPB-J1; lane 11, CFPB-K1; lane 12, CFPB-L1; lane 13, CFPB-M1; lane 14, CFPB-N1.
    Rsa I, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 194 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Fisher Scientific rsa i
    Amplified products of the 16S rDNA of various Alex-CFPB (A) and digestion of the PCR products with <t>Rsa</t> I (B), Bst UI (C), <t>Mbo</t> I (D), and Hha I (E). Lane M, 100-bp ladder markers; lane 1, CFPB-A9; lane 2, CFPB-B1; lane 3, CFPB-C1; lane 4, CFPB-D1; lane 5, CFPB-E1; lane 6, CFPB-F1; lane 7, CFPB-G1; lane 8, CFPB-H1; lane 9, CFPB-I1; lane 10, CFPB-J1; lane 11, CFPB-K1; lane 12, CFPB-L1; lane 13, CFPB-M1; lane 14, CFPB-N1.
    Rsa I, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    TA Instruments rsa 3
    Amplified products of the 16S rDNA of various Alex-CFPB (A) and digestion of the PCR products with <t>Rsa</t> I (B), Bst UI (C), <t>Mbo</t> I (D), and Hha I (E). Lane M, 100-bp ladder markers; lane 1, CFPB-A9; lane 2, CFPB-B1; lane 3, CFPB-C1; lane 4, CFPB-D1; lane 5, CFPB-E1; lane 6, CFPB-F1; lane 7, CFPB-G1; lane 8, CFPB-H1; lane 9, CFPB-I1; lane 10, CFPB-J1; lane 11, CFPB-K1; lane 12, CFPB-L1; lane 13, CFPB-M1; lane 14, CFPB-N1.
    Rsa 3, supplied by TA Instruments, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Agilent technologies rsa i
    Amplified products of the 16S rDNA of various Alex-CFPB (A) and digestion of the PCR products with <t>Rsa</t> I (B), Bst UI (C), <t>Mbo</t> I (D), and Hha I (E). Lane M, 100-bp ladder markers; lane 1, CFPB-A9; lane 2, CFPB-B1; lane 3, CFPB-C1; lane 4, CFPB-D1; lane 5, CFPB-E1; lane 6, CFPB-F1; lane 7, CFPB-G1; lane 8, CFPB-H1; lane 9, CFPB-I1; lane 10, CFPB-J1; lane 11, CFPB-K1; lane 12, CFPB-L1; lane 13, CFPB-M1; lane 14, CFPB-N1.
    Rsa I, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Allelic discrimination plots obtained for SNP103 using TaqMan, KASP and rhAmp genotyping assays on 96 samples. Red and blue dots represent the homozygous genotypes, the green circles represent heterozygous genotypes and the squares on the bottom left of the plot are no-template control

    Journal: Plant Methods

    Article Title: Comparison of three PCR-based assays for SNP genotyping in plants

    doi: 10.1186/s13007-018-0295-6

    Figure Lengend Snippet: Allelic discrimination plots obtained for SNP103 using TaqMan, KASP and rhAmp genotyping assays on 96 samples. Red and blue dots represent the homozygous genotypes, the green circles represent heterozygous genotypes and the squares on the bottom left of the plot are no-template control

    Article Snippet: KASP genotyping assay Genotyping by KASP was performed following manufacture’s instruction using 10 ng of DNA mixed with the KASP Genotyping Master Mix (Catalogue number: KBS-1016-017, LGC Genomics, Hoddesdon, UK) and custom KASP SNP assays (LGC Genomics). rhAmp genotyping assay Genotyping by rhAmp was performed following manufacture’s instruction using 5 ng of DNA mixed with rhAmp Genotyping Mix, composed by rhAmp Genotyping Master Mix (catalogue number: 1076017, Integrated DNA Technology, Coralville, Iowa, USA), rhAmp Reporter Mix (catalogue number: 1076028, Integrated DNA Technology) and custom rhAmp SNP assays (Integrated DNA Technology).

    Techniques:

    In vivo biodistribution of ABD-fused (Z HER2:342 ) 2 in a preclinical rat model. A , ELISA measurements showing pH-dependent binding of srFcRn to RSA in complex with ABD-(Z HER2:342 ) 2 . The numbers given represent the mean of triplicates. The blood, skin, and muscle biodistribution of 111 In-labeled-RSA ( B ) and 177 Lu-labeled-ABD-(Z HER2:342 ) 2 ( C ) in Sprague-Dawley rats (three rats/group and time point). Organ uptake is expressed as percent of injected radioactivity/g (% IA/g ), and error bars and corrected for interstitial volume as described under “Experimental Procedures.”

    Journal: The Journal of Biological Chemistry

    Article Title: Extending Half-life by Indirect Targeting of the Neonatal Fc Receptor (FcRn) Using a Minimal Albumin Binding Domain *

    doi: 10.1074/jbc.M110.164848

    Figure Lengend Snippet: In vivo biodistribution of ABD-fused (Z HER2:342 ) 2 in a preclinical rat model. A , ELISA measurements showing pH-dependent binding of srFcRn to RSA in complex with ABD-(Z HER2:342 ) 2 . The numbers given represent the mean of triplicates. The blood, skin, and muscle biodistribution of 111 In-labeled-RSA ( B ) and 177 Lu-labeled-ABD-(Z HER2:342 ) 2 ( C ) in Sprague-Dawley rats (three rats/group and time point). Organ uptake is expressed as percent of injected radioactivity/g (% IA/g ), and error bars and corrected for interstitial volume as described under “Experimental Procedures.”

    Article Snippet: The chelator CHX-A″/DTPA (Macrocyclics, Dallas, TX) was coupled to RSA (Sigma) or ABD-(ZHER2:342 )2 at a 1:1 molar ratio to avoid overmodification; 200 μl of RSA (5 mg/ml in 0.07 m borate buffer, pH 9.2) was mixed with 10 μl of a freshly prepared solution of CHX-A″/DTPA (1 mg/ml in 0.07 m borate buffer, pH 9.2), and 290 μl of 0.07 m borate buffer, pH 9.2, was added to the mixture.

    Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Binding Assay, Labeling, Injection, Radioactivity, IA

    ( A ) Image of 96 Rsa I-digested HFE amplicons analyzed in two sequential 48-sample separations. ( B and C ) Expanded views of the separation image from the first and second injection, respectively. The total time for the two separations was less than 7.6 min. Samples were separated on 0.75% (wt/vol) hydroxyethylcellulose in 1× TBE buffer with 1 μM ethidium bromide in the running buffer. The channels were 10 cm in length from the injection to the detection region. The injection was performed as described in the text and the applied fields for injection and separation were 300 V/cm.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: High-throughput genetic analysis using microfabricated 96-sample capillary array electrophoresis microplates

    doi:

    Figure Lengend Snippet: ( A ) Image of 96 Rsa I-digested HFE amplicons analyzed in two sequential 48-sample separations. ( B and C ) Expanded views of the separation image from the first and second injection, respectively. The total time for the two separations was less than 7.6 min. Samples were separated on 0.75% (wt/vol) hydroxyethylcellulose in 1× TBE buffer with 1 μM ethidium bromide in the running buffer. The channels were 10 cm in length from the injection to the detection region. The injection was performed as described in the text and the applied fields for injection and separation were 300 V/cm.

    Article Snippet: The restriction digestion of amplified product was carried out by adding 4 μl of each amplified sample to 6 μl of buffer containing 2 units of Rsa I (Sigma) and digesting for 90 min at 37°C.

    Techniques: Injection

    Comparison of results obtained with three RSA methods. The Percoll method is shown in black , the filtration method in gray , and the sorbitol-only method in pattern. Across the three methods, results are consistent for each line and the distinction between sensitive and surviving is clear. GB4 and progeny 39E3, 34F5, 36F11, and 24G11 all contain K13 C580, while 803 and progeny 76H10 and 61E8 contain K13 C580Y. Survival of all K13 C580 lines is significantly less than those containing the K13 C580Y polymorphism. Values are presented in detail in Additional file 2

    Journal: Malaria Journal

    Article Title: Alternative methods for the Plasmodium falciparum artemisinin ring-stage survival assay with increased simplicity and parasite stage-specificity

    doi: 10.1186/s12936-016-1148-2

    Figure Lengend Snippet: Comparison of results obtained with three RSA methods. The Percoll method is shown in black , the filtration method in gray , and the sorbitol-only method in pattern. Across the three methods, results are consistent for each line and the distinction between sensitive and surviving is clear. GB4 and progeny 39E3, 34F5, 36F11, and 24G11 all contain K13 C580, while 803 and progeny 76H10 and 61E8 contain K13 C580Y. Survival of all K13 C580 lines is significantly less than those containing the K13 C580Y polymorphism. Values are presented in detail in Additional file 2

    Article Snippet: Percoll gradient ring-stage survival assay To perform the 0–3 h RSA protocol developed by Witkowski, Amaratunga, and colleagues [ ] (henceforth designated as the Percoll gradient RSA), 300 µL of packed parasitized RBCs containing 2–3 % mature schizonts ( > 10 well-segmented nuclei) were collected from a P. falciparum culture previously synchronized twice with 5 % sorbitol (Sigma-Aldrich, MO, USA) dissolved in distilled tissue culture-grade water (Gibco, MD, USA), 10 min, 37 °C, over two parasite intra-erythrocytic cycles.

    Techniques: Filtration

    Apoptosis-related cellular events after AOPPs–RSA treatment. (A,B) Pro-caspase 9 significantly decreased from 12 h, accompanied by an overexpression of cleaved-caspase 9 in a dose-dependent manner. (C,D) Cleaved-caspase 3 increased from 12 h, however, pro-caspase 3 had no significant changes. (E) PARP-1 and cleaved-PARP-1 activation were enhanced by AOPPs–RSA at increasing concentrations. (F) PARP-1 and cleaved-PARP-1 increased from 2 h and reached the highest level at 24 h 05). Data represent mean ± SEM of at least three independent experiments. ∗ P

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Advanced Oxidative Protein Products Cause Pain Hypersensitivity in Rats by Inducing Dorsal Root Ganglion Neurons Apoptosis via NADPH Oxidase 4/c-Jun N-terminal Kinase Pathways

    doi: 10.3389/fnmol.2017.00195

    Figure Lengend Snippet: Apoptosis-related cellular events after AOPPs–RSA treatment. (A,B) Pro-caspase 9 significantly decreased from 12 h, accompanied by an overexpression of cleaved-caspase 9 in a dose-dependent manner. (C,D) Cleaved-caspase 3 increased from 12 h, however, pro-caspase 3 had no significant changes. (E) PARP-1 and cleaved-PARP-1 activation were enhanced by AOPPs–RSA at increasing concentrations. (F) PARP-1 and cleaved-PARP-1 increased from 2 h and reached the highest level at 24 h 05). Data represent mean ± SEM of at least three independent experiments. ∗ P

    Article Snippet: AOPPs–RSA (50 mg/kg), RSA (50 mg/kg), and PBS (pH 7.4) were intravenously injected each day of the study, Antioxidant apocynin (50 mg/kg, Sigma–Aldrich Corp, MO, United States) was dosed by intragastric administration each day of the study duration ( ).

    Techniques: Over Expression, Activation Assay

    Advanced oxidative protein products -induced DRG neurons apoptosis. (A) Viability of DRG neurons treated with AOPPs at different concentrations. (B) Representative dot blots of Annexin V–FITC versus PI. Primary DRG neurons were incubated with AOPPs–RSA for the indicated concentrations or unmodified RSA for 24 h. (C) DRG neurons were incubated with 100 μg/mL AOPPs–RSA for the indicated time durations. Apoptosis was quantified by measuring the number of combined early and late apoptotic cells using flow cytometry and was found to increase in a concentration and time-dependent manner. Data represent mean ± SEM of at least three independent experiments. ∗ P

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Advanced Oxidative Protein Products Cause Pain Hypersensitivity in Rats by Inducing Dorsal Root Ganglion Neurons Apoptosis via NADPH Oxidase 4/c-Jun N-terminal Kinase Pathways

    doi: 10.3389/fnmol.2017.00195

    Figure Lengend Snippet: Advanced oxidative protein products -induced DRG neurons apoptosis. (A) Viability of DRG neurons treated with AOPPs at different concentrations. (B) Representative dot blots of Annexin V–FITC versus PI. Primary DRG neurons were incubated with AOPPs–RSA for the indicated concentrations or unmodified RSA for 24 h. (C) DRG neurons were incubated with 100 μg/mL AOPPs–RSA for the indicated time durations. Apoptosis was quantified by measuring the number of combined early and late apoptotic cells using flow cytometry and was found to increase in a concentration and time-dependent manner. Data represent mean ± SEM of at least three independent experiments. ∗ P

    Article Snippet: AOPPs–RSA (50 mg/kg), RSA (50 mg/kg), and PBS (pH 7.4) were intravenously injected each day of the study, Antioxidant apocynin (50 mg/kg, Sigma–Aldrich Corp, MO, United States) was dosed by intragastric administration each day of the study duration ( ).

    Techniques: Incubation, Flow Cytometry, Cytometry, Concentration Assay

    Advanced oxidative protein products activated JNK via Nox4–ROS pathway. (A) DRG neurons were incubated in AOPPs–RSA, unmodified RSA, or control medium for 6 h prior to DCFH-DA (10 μM) treatment. (B) DRG neurons were incubated in 200 μg/mL AOPPs–RSA for indicated time durations followed by DCFH-DA treatment. (C) DRG neurons were treated with AOPPs–RSA (200 μg/mL) with or without Nox4 siRNA, DPI (10 μM, 2 h) or NAC (2 mM, 2 h). (D) DRG neurons were cultured with control medium, RSA, AOPPs–RSA (200 μg/mL), AOPPs+siNox4, AOPPs+DPI, or AOPPs+NAC for 360 min. Confocal laser scanning microscope system was used to visualize ROS generation in DRG neurons with the use of DCFH-DA, scale bar, 30 μm. (E,F) The level of Nox4 was up-regulated significantly in DRG neurons incubated with AOPPs–RSA in indicated concentrations or periods. (G,H) The expression level of JNK/p-JNK. Data represent mean ± SEM of at least three independent experiments. ∗ P

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Advanced Oxidative Protein Products Cause Pain Hypersensitivity in Rats by Inducing Dorsal Root Ganglion Neurons Apoptosis via NADPH Oxidase 4/c-Jun N-terminal Kinase Pathways

    doi: 10.3389/fnmol.2017.00195

    Figure Lengend Snippet: Advanced oxidative protein products activated JNK via Nox4–ROS pathway. (A) DRG neurons were incubated in AOPPs–RSA, unmodified RSA, or control medium for 6 h prior to DCFH-DA (10 μM) treatment. (B) DRG neurons were incubated in 200 μg/mL AOPPs–RSA for indicated time durations followed by DCFH-DA treatment. (C) DRG neurons were treated with AOPPs–RSA (200 μg/mL) with or without Nox4 siRNA, DPI (10 μM, 2 h) or NAC (2 mM, 2 h). (D) DRG neurons were cultured with control medium, RSA, AOPPs–RSA (200 μg/mL), AOPPs+siNox4, AOPPs+DPI, or AOPPs+NAC for 360 min. Confocal laser scanning microscope system was used to visualize ROS generation in DRG neurons with the use of DCFH-DA, scale bar, 30 μm. (E,F) The level of Nox4 was up-regulated significantly in DRG neurons incubated with AOPPs–RSA in indicated concentrations or periods. (G,H) The expression level of JNK/p-JNK. Data represent mean ± SEM of at least three independent experiments. ∗ P

    Article Snippet: AOPPs–RSA (50 mg/kg), RSA (50 mg/kg), and PBS (pH 7.4) were intravenously injected each day of the study, Antioxidant apocynin (50 mg/kg, Sigma–Aldrich Corp, MO, United States) was dosed by intragastric administration each day of the study duration ( ).

    Techniques: Incubation, Cell Culture, Laser-Scanning Microscopy, Expressing

    Advanced oxidative protein products–RSA induced mitochondrial dysfunction in primary DRG neurons. (A) The mitochondrial membrane potential (MMP) was observed in AOPPs-treated DRG neurons. The cells were incubated with 100 μg/mL AOPPs–RSA for 24 h and then JC-1 dye for 20 min. The cells were observed using a confocal laser scanning microscope system and demonstrated an AOPPs-dependent MMP loss in a dose-dependent manner, scale bar, 20 μm. (B–G) Proapoptotic Bax increased and antiapoptotic Bcl-2 decreased from 12 h; prolonged incubation and a higher concentration of AOPPs–RSA increased the Bax/Bcl-2 ratio. (H,I) Cytochrome c increased in AOPPs-treated DRg neurons. Data represent mean ± SEM of at least 3 independent experiments. ∗ P

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Advanced Oxidative Protein Products Cause Pain Hypersensitivity in Rats by Inducing Dorsal Root Ganglion Neurons Apoptosis via NADPH Oxidase 4/c-Jun N-terminal Kinase Pathways

    doi: 10.3389/fnmol.2017.00195

    Figure Lengend Snippet: Advanced oxidative protein products–RSA induced mitochondrial dysfunction in primary DRG neurons. (A) The mitochondrial membrane potential (MMP) was observed in AOPPs-treated DRG neurons. The cells were incubated with 100 μg/mL AOPPs–RSA for 24 h and then JC-1 dye for 20 min. The cells were observed using a confocal laser scanning microscope system and demonstrated an AOPPs-dependent MMP loss in a dose-dependent manner, scale bar, 20 μm. (B–G) Proapoptotic Bax increased and antiapoptotic Bcl-2 decreased from 12 h; prolonged incubation and a higher concentration of AOPPs–RSA increased the Bax/Bcl-2 ratio. (H,I) Cytochrome c increased in AOPPs-treated DRg neurons. Data represent mean ± SEM of at least 3 independent experiments. ∗ P

    Article Snippet: AOPPs–RSA (50 mg/kg), RSA (50 mg/kg), and PBS (pH 7.4) were intravenously injected each day of the study, Antioxidant apocynin (50 mg/kg, Sigma–Aldrich Corp, MO, United States) was dosed by intragastric administration each day of the study duration ( ).

    Techniques: Incubation, Laser-Scanning Microscopy, Concentration Assay

    Advanced oxidative protein products-induced DRG neurons apoptosis was mediated through the Nox4–JNK–caspase cascade–PARP pathway. (A) DRG neurons were pre-incubated with siRNA Nox4, DPI (10 μM), NAC (2 mM), an JNK inhibitor (SP600125, 10 μM) or the broad-spectrum caspase inhibitor Z-VAD-fmk (20 μM) before AOPPs–RSA treatment. They all inhibited AOPPs-induced cell apoptosis. (B) The expression of PARP-1/ cleaved-PARP-1 were tested by western blots. Data represent mean ± SEM of at least three independent experiments. ∗ P

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Advanced Oxidative Protein Products Cause Pain Hypersensitivity in Rats by Inducing Dorsal Root Ganglion Neurons Apoptosis via NADPH Oxidase 4/c-Jun N-terminal Kinase Pathways

    doi: 10.3389/fnmol.2017.00195

    Figure Lengend Snippet: Advanced oxidative protein products-induced DRG neurons apoptosis was mediated through the Nox4–JNK–caspase cascade–PARP pathway. (A) DRG neurons were pre-incubated with siRNA Nox4, DPI (10 μM), NAC (2 mM), an JNK inhibitor (SP600125, 10 μM) or the broad-spectrum caspase inhibitor Z-VAD-fmk (20 μM) before AOPPs–RSA treatment. They all inhibited AOPPs-induced cell apoptosis. (B) The expression of PARP-1/ cleaved-PARP-1 were tested by western blots. Data represent mean ± SEM of at least three independent experiments. ∗ P

    Article Snippet: AOPPs–RSA (50 mg/kg), RSA (50 mg/kg), and PBS (pH 7.4) were intravenously injected each day of the study, Antioxidant apocynin (50 mg/kg, Sigma–Aldrich Corp, MO, United States) was dosed by intragastric administration each day of the study duration ( ).

    Techniques: Incubation, Expressing, Western Blot

    MMP-2 activity, AGE accumulation and RAGE expression in the renal tubules of diabetic or AGE-RSA-infused rats. (A) MMP-2 activity in 32-week Ctrl or diabetic renal tubules with or without ramipril treatment was determined by gelatin zymography ( n = 4). (B) AGE protein levels in 32-week Ctrl or diabetic renal tubules with or without ramipril treatment were evaluated by enzyme-linked immunosorbent assay for carboxymethyllisine ( n = 7). (C) RAGE protein expression in 32-week Ctrl or diabetic renal tubules with or without ramipril treatment was determined by western blot. Data were normalized by the intensity of α-tubulin bands. ( n = 4). (D) RAGE gene expression in total kidney cortex of AGE-RSA- or RSA-infused rats was evaluated by quantitative real-time RT-PCR (n = 7). (E) MMP-2 activity in the tubules of AGE-RSA- or RSA-infused rats was determined by gelatin zymography (n = 4). Data are shown as mean ± SEM.

    Journal: Diabetology & Metabolic Syndrome

    Article Title: Ramipril inhibits AGE-RAGE-induced matrix metalloproteinase-2 activation in experimental diabetic nephropathy

    doi: 10.1186/1758-5996-6-86

    Figure Lengend Snippet: MMP-2 activity, AGE accumulation and RAGE expression in the renal tubules of diabetic or AGE-RSA-infused rats. (A) MMP-2 activity in 32-week Ctrl or diabetic renal tubules with or without ramipril treatment was determined by gelatin zymography ( n = 4). (B) AGE protein levels in 32-week Ctrl or diabetic renal tubules with or without ramipril treatment were evaluated by enzyme-linked immunosorbent assay for carboxymethyllisine ( n = 7). (C) RAGE protein expression in 32-week Ctrl or diabetic renal tubules with or without ramipril treatment was determined by western blot. Data were normalized by the intensity of α-tubulin bands. ( n = 4). (D) RAGE gene expression in total kidney cortex of AGE-RSA- or RSA-infused rats was evaluated by quantitative real-time RT-PCR (n = 7). (E) MMP-2 activity in the tubules of AGE-RSA- or RSA-infused rats was determined by gelatin zymography (n = 4). Data are shown as mean ± SEM.

    Article Snippet: Preparation of AGE-RSA and AGE-modified bovine serum albumin (AGE-BSA) AGE-RSA and AGE-BSA were prepared by incubating RSA or BSA (Fraction V, Sigma Chemical Co, St. Louis, MO, USA) with 0.5 M D-glucose in PBS at 37°C for 3 months as previously described [ ].

    Techniques: Activity Assay, Expressing, Zymography, Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR

    Boxplots with IgG and IgM anti- P. jirovecii levels results across patients with PcP and patients without P. jirovecii infection. Simple boxplots showing the reactivity levels (OD at 405 nm) of IgG and IgM anti- P. jirovecii antibodies detected by ELISA protocols applied with the Kex1 RSA (A) and Msg RSA (B) as coating antigens, in all sera specimens of patients analyzed in the study. The statistic values ( p ) representing the statistical significant difference from Mann Whitney-U tests performed between patient’s groups are presented.

    Journal: Frontiers in Microbiology

    Article Title: Development of a Gold Nanoparticle-Based Lateral-Flow Immunoassay for Pneumocystis Pneumonia Serological Diagnosis at Point-of-Care

    doi: 10.3389/fmicb.2019.02917

    Figure Lengend Snippet: Boxplots with IgG and IgM anti- P. jirovecii levels results across patients with PcP and patients without P. jirovecii infection. Simple boxplots showing the reactivity levels (OD at 405 nm) of IgG and IgM anti- P. jirovecii antibodies detected by ELISA protocols applied with the Kex1 RSA (A) and Msg RSA (B) as coating antigens, in all sera specimens of patients analyzed in the study. The statistic values ( p ) representing the statistical significant difference from Mann Whitney-U tests performed between patient’s groups are presented.

    Article Snippet: The control and test lines were spotted manually, in a circle or in a line, by depositing 0.03 mg.mL–1 of anti-RSA antibodies (anti-Msg or anti-Kex1, depending on the RSA present in the conjugate) and 0.001 mg.mL–1 of anti-human IgM antibodies (I-0759, Sigma® ), respectively.

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Assessment of the RSA expression and purification process. SDS-PAGE profiles of non-induced (1) and induced (2) E. coli XJb (DE3) cells transformed with the expression vector with the Kex1 RSA (A) and the Msg RSA (B) ; and SDS-PAGE profiles of purification products of the Msg RSA and Kex1 RSA, after IMAC protocol (C) . Two different protein molecular weight standards were used, the Roti § -Mark standard from Carl-Roth § (M1) and one produced “in house” (M2). The bands of interest and their theoretical molecular weight are highlighted (16.7 kDa for Msg RSA and 22 kDa for Kex1 RSA).

    Journal: Frontiers in Microbiology

    Article Title: Development of a Gold Nanoparticle-Based Lateral-Flow Immunoassay for Pneumocystis Pneumonia Serological Diagnosis at Point-of-Care

    doi: 10.3389/fmicb.2019.02917

    Figure Lengend Snippet: Assessment of the RSA expression and purification process. SDS-PAGE profiles of non-induced (1) and induced (2) E. coli XJb (DE3) cells transformed with the expression vector with the Kex1 RSA (A) and the Msg RSA (B) ; and SDS-PAGE profiles of purification products of the Msg RSA and Kex1 RSA, after IMAC protocol (C) . Two different protein molecular weight standards were used, the Roti § -Mark standard from Carl-Roth § (M1) and one produced “in house” (M2). The bands of interest and their theoretical molecular weight are highlighted (16.7 kDa for Msg RSA and 22 kDa for Kex1 RSA).

    Article Snippet: The control and test lines were spotted manually, in a circle or in a line, by depositing 0.03 mg.mL–1 of anti-RSA antibodies (anti-Msg or anti-Kex1, depending on the RSA present in the conjugate) and 0.001 mg.mL–1 of anti-human IgM antibodies (I-0759, Sigma® ), respectively.

    Techniques: Expressing, Purification, SDS Page, Transformation Assay, Plasmid Preparation, Molecular Weight, Produced

    Interaction of AuNP-Msg-Casein (A,B) and AuNP-Kex1-Casein conjugates (C,D) with sera pools from patients with and without P. jirovecii infection. (A,C) Agarose gel electrophoresis of AuNP-RSA-Casein conjugates before interaction with serum (1) and after interaction with positive (2) and negative (3) serum samples. (B) Electrophoretic mobility of the AuNP-RSA-Casein conjugates before interaction with human sera (No Serum) and after interaction with the positive human sera (PosSerum) and negative human sera (NegSerum). Shifts (μm.cm/V.s) between blocked conjugates with positive and with negative sera are indicated, evidencing the binding of antibodies present in the positive serum. Standard deviation bars correspond to triplicate experiments.

    Journal: Frontiers in Microbiology

    Article Title: Development of a Gold Nanoparticle-Based Lateral-Flow Immunoassay for Pneumocystis Pneumonia Serological Diagnosis at Point-of-Care

    doi: 10.3389/fmicb.2019.02917

    Figure Lengend Snippet: Interaction of AuNP-Msg-Casein (A,B) and AuNP-Kex1-Casein conjugates (C,D) with sera pools from patients with and without P. jirovecii infection. (A,C) Agarose gel electrophoresis of AuNP-RSA-Casein conjugates before interaction with serum (1) and after interaction with positive (2) and negative (3) serum samples. (B) Electrophoretic mobility of the AuNP-RSA-Casein conjugates before interaction with human sera (No Serum) and after interaction with the positive human sera (PosSerum) and negative human sera (NegSerum). Shifts (μm.cm/V.s) between blocked conjugates with positive and with negative sera are indicated, evidencing the binding of antibodies present in the positive serum. Standard deviation bars correspond to triplicate experiments.

    Article Snippet: The control and test lines were spotted manually, in a circle or in a line, by depositing 0.03 mg.mL–1 of anti-RSA antibodies (anti-Msg or anti-Kex1, depending on the RSA present in the conjugate) and 0.001 mg.mL–1 of anti-human IgM antibodies (I-0759, Sigma® ), respectively.

    Techniques: Infection, Agarose Gel Electrophoresis, Binding Assay, Standard Deviation

    AuNP-RSA conjugates formation. Agarose gel electrophoresis at increasing ratios of Msg RSA to AuNPs (A) and Kex1 RSA to AuNPs (C) , showing the migration of each conjugate bands toward the positive electrode. Differences between the electrophoretic mobility of the AuNPs alone and AuNP-Msg (B) or AuNP-Kex1 (D) conjugates at increasing ratios, calculated from the migration distances in gel. The nine ratios (162.5–5000) correspond to protein concentrations of 9.75, 15, 19.5, 39, 58.5, 117, 175.5, 234, and 300 nM. Standard deviation bars correspond to duplicate experiments.

    Journal: Frontiers in Microbiology

    Article Title: Development of a Gold Nanoparticle-Based Lateral-Flow Immunoassay for Pneumocystis Pneumonia Serological Diagnosis at Point-of-Care

    doi: 10.3389/fmicb.2019.02917

    Figure Lengend Snippet: AuNP-RSA conjugates formation. Agarose gel electrophoresis at increasing ratios of Msg RSA to AuNPs (A) and Kex1 RSA to AuNPs (C) , showing the migration of each conjugate bands toward the positive electrode. Differences between the electrophoretic mobility of the AuNPs alone and AuNP-Msg (B) or AuNP-Kex1 (D) conjugates at increasing ratios, calculated from the migration distances in gel. The nine ratios (162.5–5000) correspond to protein concentrations of 9.75, 15, 19.5, 39, 58.5, 117, 175.5, 234, and 300 nM. Standard deviation bars correspond to duplicate experiments.

    Article Snippet: The control and test lines were spotted manually, in a circle or in a line, by depositing 0.03 mg.mL–1 of anti-RSA antibodies (anti-Msg or anti-Kex1, depending on the RSA present in the conjugate) and 0.001 mg.mL–1 of anti-human IgM antibodies (I-0759, Sigma® ), respectively.

    Techniques: Agarose Gel Electrophoresis, Migration, Standard Deviation

    Neighbour-joining similarity tree for 16S rRNA gene sequences of most mycobacterial species. Legend: N. asteroides ATCC 49872 (Genbank Z82229) was used as the outgroup. Table 1 lists the GenBank accession numbers of the sequences used to construct this tree. ARDRA patterns for Hha I, Mbo I, Rsa I and Bst UI are listed after the species name. a: GenBank AF028712. Erroneously listed in GenBank as M. peregrinum (see also legend of Table 1 ). b: M. gastri clusters below 100% with M. kansasii, although it is generally agreed that the 16S rRNA gene sequences for M. kansasii and M. gastri are identical. This can be explained by the fact that the only available GenBank M. gastri sequence (X52919) contained several ambiguities. c. M. lentiflavum, initially not included in the manuscript is not presented in this tree. It clusters close to the branch including M. heidelbergense, M. simiae, M. triplex and M. genavense.

    Journal: BMC Microbiology

    Article Title: Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory

    doi: 10.1186/1471-2180-2-4

    Figure Lengend Snippet: Neighbour-joining similarity tree for 16S rRNA gene sequences of most mycobacterial species. Legend: N. asteroides ATCC 49872 (Genbank Z82229) was used as the outgroup. Table 1 lists the GenBank accession numbers of the sequences used to construct this tree. ARDRA patterns for Hha I, Mbo I, Rsa I and Bst UI are listed after the species name. a: GenBank AF028712. Erroneously listed in GenBank as M. peregrinum (see also legend of Table 1 ). b: M. gastri clusters below 100% with M. kansasii, although it is generally agreed that the 16S rRNA gene sequences for M. kansasii and M. gastri are identical. This can be explained by the fact that the only available GenBank M. gastri sequence (X52919) contained several ambiguities. c. M. lentiflavum, initially not included in the manuscript is not presented in this tree. It clusters close to the branch including M. heidelbergense, M. simiae, M. triplex and M. genavense.

    Article Snippet: Restriction digestion The restriction enzymes used were Hha I (isoschizomer of CfoI)(Amersham Pharmacia Biotech Benelux, Roosendaal, the Netherlands), Mbo I (Fermentas, Vilnius, Lithuania), Rsa I (Amersham Pharmacia).

    Techniques: Construct, Sequencing

    Rsa I restriction patterns of amplified mycobacterial 16S rRNA genes. Legend: M: marker (100 base pair ladder, Fermentas, Vilnius, Lithuania)

    Journal: BMC Microbiology

    Article Title: Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory

    doi: 10.1186/1471-2180-2-4

    Figure Lengend Snippet: Rsa I restriction patterns of amplified mycobacterial 16S rRNA genes. Legend: M: marker (100 base pair ladder, Fermentas, Vilnius, Lithuania)

    Article Snippet: Restriction digestion The restriction enzymes used were Hha I (isoschizomer of CfoI)(Amersham Pharmacia Biotech Benelux, Roosendaal, the Netherlands), Mbo I (Fermentas, Vilnius, Lithuania), Rsa I (Amersham Pharmacia).

    Techniques: Amplification, Marker

    Rsa I restriction patterns of mycobacterial 16S rRNA genes, theoretically calculated using RFLP (Applied Maths) and published GenBank sequences. Graphical representation and table of restriction fragment lengths for each of the possible patterns.

    Journal: BMC Microbiology

    Article Title: Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of cultured mycobacteria in a diagnostic laboratory

    doi: 10.1186/1471-2180-2-4

    Figure Lengend Snippet: Rsa I restriction patterns of mycobacterial 16S rRNA genes, theoretically calculated using RFLP (Applied Maths) and published GenBank sequences. Graphical representation and table of restriction fragment lengths for each of the possible patterns.

    Article Snippet: Restriction digestion The restriction enzymes used were Hha I (isoschizomer of CfoI)(Amersham Pharmacia Biotech Benelux, Roosendaal, the Netherlands), Mbo I (Fermentas, Vilnius, Lithuania), Rsa I (Amersham Pharmacia).

    Techniques:

    Representative diversity revealed by M46 PCR-RFLP in M. catarrhalis . (A) Amplimer B; (B) amplimer C. Samples were treated with Hae III (lanes 1 to 7) or with Rsa I (lanes 8 to 14) or are undigested amplimers (lanes 15 to 21). Each lane contained 10 μl of PCR product. Lanes 1, 8, and 15, sample ATCC 25238; lanes 2, 9, and 16, sample TN16a; lanes 3, 10, and 17, sample BL2; lanes 4, 11, and 18, sample NY1a; lanes 5, 12, and 19, sample TN17a; lanes 6, 13, and 20, sample TN7a; lanes 7, 14, and 21, sample TN13a. Lanes M are 100-to-1,500-bp ladders, with double-bright bands at 600 bp and additional fragments of 2,000 bp.

    Journal: Journal of Clinical Microbiology

    Article Title: Genetic Diversity among Strains of Moraxella catarrhalis: Analysis Using Multiple DNA Probes and a Single-Locus PCR-Restriction Fragment Length Polymorphism Method

    doi:

    Figure Lengend Snippet: Representative diversity revealed by M46 PCR-RFLP in M. catarrhalis . (A) Amplimer B; (B) amplimer C. Samples were treated with Hae III (lanes 1 to 7) or with Rsa I (lanes 8 to 14) or are undigested amplimers (lanes 15 to 21). Each lane contained 10 μl of PCR product. Lanes 1, 8, and 15, sample ATCC 25238; lanes 2, 9, and 16, sample TN16a; lanes 3, 10, and 17, sample BL2; lanes 4, 11, and 18, sample NY1a; lanes 5, 12, and 19, sample TN17a; lanes 6, 13, and 20, sample TN7a; lanes 7, 14, and 21, sample TN13a. Lanes M are 100-to-1,500-bp ladders, with double-bright bands at 600 bp and additional fragments of 2,000 bp.

    Article Snippet: To detect RFLP variation within PCR-amplified regions encompassed by M46 DNA, 10-μl aliquots of PCR products were subjected to separate restriction enzyme digestion with Hae III and Rsa I (Promega Corp.) according to the manufacturer’s instructions.

    Techniques: Polymerase Chain Reaction

    Relationship between mean telomere length, measured by Southern blots of the TRFs, versus the ratio of T (telomere amount)/Dx (DNA amount), measured by dot blots, and versus the ratio of T (telomere product)/S(single gene product), measured by qPCR. ( a and b ) The TRF products generated by Hinf I/ Rsa I and by Hph I/ Mnl I versus the T/Dx. ( c and d ) The TRF product generated by Hinf I/ Rsa I and by Hph I/ Mnl I versus T/S.

    Journal: Nucleic Acids Research

    Article Title: Measurement of telomere DNA content by dot blot analysis

    doi: 10.1093/nar/gkr235

    Figure Lengend Snippet: Relationship between mean telomere length, measured by Southern blots of the TRFs, versus the ratio of T (telomere amount)/Dx (DNA amount), measured by dot blots, and versus the ratio of T (telomere product)/S(single gene product), measured by qPCR. ( a and b ) The TRF products generated by Hinf I/ Rsa I and by Hph I/ Mnl I versus the T/Dx. ( c and d ) The TRF product generated by Hinf I/ Rsa I and by Hph I/ Mnl I versus T/S.

    Article Snippet: Thereafter, samples were digested with restriction enzymes Hinf I (10 U) and Rsa I (10 U; Roche) or HphI (3.1 U)/MnlI (3.1 U) (New England Biolabs, Ipswich, MA, USA).

    Techniques: Real-time Polymerase Chain Reaction, Generated

    Relationship between mean telomere length, measured by Southern blots of the TRFs, generated by Hinf I/ Rsa I, versus the ratio of T (telomere amount)/Dx (DNA amount), measured by dot blots. Data displayed in the figure are a composite of data displayed in Figure 2 plus an additional set of measurements in leukocytes from newborns and exceptionally old persons.

    Journal: Nucleic Acids Research

    Article Title: Measurement of telomere DNA content by dot blot analysis

    doi: 10.1093/nar/gkr235

    Figure Lengend Snippet: Relationship between mean telomere length, measured by Southern blots of the TRFs, generated by Hinf I/ Rsa I, versus the ratio of T (telomere amount)/Dx (DNA amount), measured by dot blots. Data displayed in the figure are a composite of data displayed in Figure 2 plus an additional set of measurements in leukocytes from newborns and exceptionally old persons.

    Article Snippet: Thereafter, samples were digested with restriction enzymes Hinf I (10 U) and Rsa I (10 U; Roche) or HphI (3.1 U)/MnlI (3.1 U) (New England Biolabs, Ipswich, MA, USA).

    Techniques: Generated

    A PCR-RFLP assay of the ITS2 region applied to pearls from Pinctada margaritifera , P. maxima and P. radiata . (A) PCR products of 575 bp ( P. margaritifera ), 571 bp ( P. maxima ) and 590–91 bp ( P. radiata ) obtained with ITS2 universal primers (5.8S-F and 28S-R) and (B) RFLP patterns of ITS2 amplicons (from A) obtained after digestion with Rsa I. MW: molecular weight size marker, 100-bp DNA ladder; lanes 1–3: P. maxima (PMX) pearls; lane 4: P. margaritifera (PMR) pearl; lanes 5–10: P. radiata (PR) pearls; lanes 11–16: P. margaritifera pearls; lane 17: PCR negative control; lanes 18 and 19: P. radiata and P. margaritifera positive controls. Note: The P. maxima positive control is shown in Figure 4 .

    Journal: PLoS ONE

    Article Title: DNA Fingerprinting of Pearls to Determine Their Origins

    doi: 10.1371/journal.pone.0075606

    Figure Lengend Snippet: A PCR-RFLP assay of the ITS2 region applied to pearls from Pinctada margaritifera , P. maxima and P. radiata . (A) PCR products of 575 bp ( P. margaritifera ), 571 bp ( P. maxima ) and 590–91 bp ( P. radiata ) obtained with ITS2 universal primers (5.8S-F and 28S-R) and (B) RFLP patterns of ITS2 amplicons (from A) obtained after digestion with Rsa I. MW: molecular weight size marker, 100-bp DNA ladder; lanes 1–3: P. maxima (PMX) pearls; lane 4: P. margaritifera (PMR) pearl; lanes 5–10: P. radiata (PR) pearls; lanes 11–16: P. margaritifera pearls; lane 17: PCR negative control; lanes 18 and 19: P. radiata and P. margaritifera positive controls. Note: The P. maxima positive control is shown in Figure 4 .

    Article Snippet: Restriction analysis was done in 12 µl reaction mixtures with 5 µl of amplified product, 100 µg/ml bovine serum albumin (New England Biolabs, Inc.), 1.2 µl enzyme buffer (New England Biolabs, Inc.) and 0.5 units of Rsa I (Fermentas GmbH).

    Techniques: Polymerase Chain Reaction, RFLP Assay, Molecular Weight, Marker, Negative Control, Positive Control

    Blind PCR-RFLP assay with eighteen pearls of unknown identity. (A) PCR products of 575 bp ( Pinctada margaritifera ), 571 bp ( P. maxima ) and 590–91 bp ( P. radiata ) obtained with ITS2 universal primers (5.8S-F and 28S-R) and (B) of 335 bp obtained with 28S-R and the P. margaritifera specific primer ITS2-Marg-F. (C) RFLP patterns of ITS2 gene fragments (from A) obtained after digestion with Rsa I. MW: molecular weight size marker, 100 bp DNA ladder; lanes 1–18: pearl isolates; lanes 19–20: DNA extraction negative controls; lane 21: PCR negative control; lanes 22–23: P. radiata and P. margaritifera positive controls; lanes 24–26: P. radiata , P. margaritifera and P. maxima positive controls showing ITS2 PCR products (upper gel) and ITS2-RFLP patterns (lower gel).

    Journal: PLoS ONE

    Article Title: DNA Fingerprinting of Pearls to Determine Their Origins

    doi: 10.1371/journal.pone.0075606

    Figure Lengend Snippet: Blind PCR-RFLP assay with eighteen pearls of unknown identity. (A) PCR products of 575 bp ( Pinctada margaritifera ), 571 bp ( P. maxima ) and 590–91 bp ( P. radiata ) obtained with ITS2 universal primers (5.8S-F and 28S-R) and (B) of 335 bp obtained with 28S-R and the P. margaritifera specific primer ITS2-Marg-F. (C) RFLP patterns of ITS2 gene fragments (from A) obtained after digestion with Rsa I. MW: molecular weight size marker, 100 bp DNA ladder; lanes 1–18: pearl isolates; lanes 19–20: DNA extraction negative controls; lane 21: PCR negative control; lanes 22–23: P. radiata and P. margaritifera positive controls; lanes 24–26: P. radiata , P. margaritifera and P. maxima positive controls showing ITS2 PCR products (upper gel) and ITS2-RFLP patterns (lower gel).

    Article Snippet: Restriction analysis was done in 12 µl reaction mixtures with 5 µl of amplified product, 100 µg/ml bovine serum albumin (New England Biolabs, Inc.), 1.2 µl enzyme buffer (New England Biolabs, Inc.) and 0.5 units of Rsa I (Fermentas GmbH).

    Techniques: Polymerase Chain Reaction, RFLP Assay, Molecular Weight, Marker, DNA Extraction, Negative Control

    ( a ) Western blot analysis of PC1 and PC2 protein expression in human islets cultured for 48 hours in the presence or absence of IL-1β (50 U/mL), IFN-γ (1,000 U/mL), or IL-1β plus IFN-γ. ( b ) Competitive PCR analysis of PC1 and PC2 mRNA expression in control and cytokine-cultured (IL-1β + IFN-γ, 48 hours) human islets. Lanes 1–4 contain the same amount of cDNA from the same human islet preparation and a decreasing amount of competitor DNA (0.5, 0.125, 0.03, and 0.0075 amol, respectively). Competitor DNA for PC2 was prepared from rat cDNA for PC1 from human cDNA. After amplification, PC1 products were directly run on 2% agarose-ethidium bromide gel, whereas PC2 products were digested with Rsa I before separation. Competitor rat PC2 appears as 2 bands (151 and 65 bp) after Rsa I restriction, but human PC2 remains intact. The figures are representative of 3 independent experiments.

    Journal: Journal of Clinical Investigation

    Article Title: Exposure of human islets to cytokines can result in disproportionately elevated proinsulin release

    doi:

    Figure Lengend Snippet: ( a ) Western blot analysis of PC1 and PC2 protein expression in human islets cultured for 48 hours in the presence or absence of IL-1β (50 U/mL), IFN-γ (1,000 U/mL), or IL-1β plus IFN-γ. ( b ) Competitive PCR analysis of PC1 and PC2 mRNA expression in control and cytokine-cultured (IL-1β + IFN-γ, 48 hours) human islets. Lanes 1–4 contain the same amount of cDNA from the same human islet preparation and a decreasing amount of competitor DNA (0.5, 0.125, 0.03, and 0.0075 amol, respectively). Competitor DNA for PC2 was prepared from rat cDNA for PC1 from human cDNA. After amplification, PC1 products were directly run on 2% agarose-ethidium bromide gel, whereas PC2 products were digested with Rsa I before separation. Competitor rat PC2 appears as 2 bands (151 and 65 bp) after Rsa I restriction, but human PC2 remains intact. The figures are representative of 3 independent experiments.

    Article Snippet: To discriminate the human and rat PC2 PCR fragments, 12 μL of each PCR reaction was digested overnight at 37°C with 15 U of Rsa I (Eurogentec, Seraing, Belgium) before loading on the gel.

    Techniques: Western Blot, Expressing, Cell Culture, Polymerase Chain Reaction, Amplification

    Amplified products of the 16S rDNA of various Alex-CFPB (A) and digestion of the PCR products with Rsa I (B), Bst UI (C), Mbo I (D), and Hha I (E). Lane M, 100-bp ladder markers; lane 1, CFPB-A9; lane 2, CFPB-B1; lane 3, CFPB-C1; lane 4, CFPB-D1; lane 5, CFPB-E1; lane 6, CFPB-F1; lane 7, CFPB-G1; lane 8, CFPB-H1; lane 9, CFPB-I1; lane 10, CFPB-J1; lane 11, CFPB-K1; lane 12, CFPB-L1; lane 13, CFPB-M1; lane 14, CFPB-N1.

    Journal: Applied and Environmental Microbiology

    Article Title: Population Structure of Alexandrium (Dinophyceae) Cyst Formation-Promoting Bacteria in Hiroshima Bay, Japan

    doi: 10.1128/AEM.69.11.6560-6568.2003

    Figure Lengend Snippet: Amplified products of the 16S rDNA of various Alex-CFPB (A) and digestion of the PCR products with Rsa I (B), Bst UI (C), Mbo I (D), and Hha I (E). Lane M, 100-bp ladder markers; lane 1, CFPB-A9; lane 2, CFPB-B1; lane 3, CFPB-C1; lane 4, CFPB-D1; lane 5, CFPB-E1; lane 6, CFPB-F1; lane 7, CFPB-G1; lane 8, CFPB-H1; lane 9, CFPB-I1; lane 10, CFPB-J1; lane 11, CFPB-K1; lane 12, CFPB-L1; lane 13, CFPB-M1; lane 14, CFPB-N1.

    Article Snippet: Two micrograms of the DNA was then digested with one of four restriction endonucleases, Mbo I, Hha I (TaKaRa Biomedicals, Osaka, Japan), Rsa I (Toyobo, Osaka, Japan), and Bst UI (New England Biolabs, Inc., Beverly, Mass.) according to the protocols of each manufacturer.

    Techniques: Amplification, Polymerase Chain Reaction