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  • 94
    tiangen biotech co gdna eraser
    <t>DNA</t> identification and gene transcripts analysis . A) <t>PCR</t> analyses of transgenic hairy root lines. Analyses for the presence of AaPMT (1269 bp) and AaTRI genes (1074 bp) in PT lines (1), AaPMT gene in P lines (2) and AaTRI gene in T lines (3), respectively. M, DL-2000 Marker. PC, positive control (vector contains corresponding gene). BC, blank control (hairy root generate from blank-vector transformation). B) Effects of transformation on the expression of related genes. AaPMT T and AaTRI T represent total gene of AaPMT and AaTRI respectively, AaPMT E and AaTRI E represent endogenous AaPMT and AaTRI respectively. C) Real-time fluorescence quantitative PCR analysis of the expressions of AaPMT and AaTRI in transgenic hairy roots.
    Gdna Eraser, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 94/100, based on 428 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co gdna purification kit
    Identification of S-layer proteins of L. <t>salivarius</t> REN. ( A ) SDS-PAGE of cell surface proteins extracted from L. salivarius REN. ( B ) Gene organization of the region surrounding the cbpA and nam - amidase locus. Arrows indicate the locations of primers used for PCR analysis. The red shaded regions represented the deletion target. The green and blue shaded regions represented the upstream and downstream fragments in target genes. The chromosomal map is not drawn to scale. ( C ) Schematic overview of the construction of Δ cbpA and Δ nam - amidase mutants. ( D ) PCR identification of double crossover recombinant. Lane M: <t>DNA</t> marker DL 2000, lane 1: PCR amplification of cbpA in Δ cbpA mutants, lane 2: PCR amplification of cbpA in wild type strain, lane 3: PCR amplification of nam - amidase in wild type strain, lane 4: PCR amplification of nam - amidase in Δ nam - amidase mutants. ( E ) Adhesion of L. salivarius REN, Δ cbpA and Δ nam - amidase mutants to the HT-29 cells. Data were analyzed by the paired Student’s t test (* P
    Gdna Purification Kit, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 83/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co gdna dispelling rt supermix
    Identification of S-layer proteins of L. <t>salivarius</t> REN. ( A ) SDS-PAGE of cell surface proteins extracted from L. salivarius REN. ( B ) Gene organization of the region surrounding the cbpA and nam - amidase locus. Arrows indicate the locations of primers used for PCR analysis. The red shaded regions represented the deletion target. The green and blue shaded regions represented the upstream and downstream fragments in target genes. The chromosomal map is not drawn to scale. ( C ) Schematic overview of the construction of Δ cbpA and Δ nam - amidase mutants. ( D ) PCR identification of double crossover recombinant. Lane M: <t>DNA</t> marker DL 2000, lane 1: PCR amplification of cbpA in Δ cbpA mutants, lane 2: PCR amplification of cbpA in wild type strain, lane 3: PCR amplification of nam - amidase in wild type strain, lane 4: PCR amplification of nam - amidase in Δ nam - amidase mutants. ( E ) Adhesion of L. salivarius REN, Δ cbpA and Δ nam - amidase mutants to the HT-29 cells. Data were analyzed by the paired Student’s t test (* P
    Gdna Dispelling Rt Supermix, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 94/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co fastking gdna dispelling rt supermix
    Identification of S-layer proteins of L. <t>salivarius</t> REN. ( A ) SDS-PAGE of cell surface proteins extracted from L. salivarius REN. ( B ) Gene organization of the region surrounding the cbpA and nam - amidase locus. Arrows indicate the locations of primers used for PCR analysis. The red shaded regions represented the deletion target. The green and blue shaded regions represented the upstream and downstream fragments in target genes. The chromosomal map is not drawn to scale. ( C ) Schematic overview of the construction of Δ cbpA and Δ nam - amidase mutants. ( D ) PCR identification of double crossover recombinant. Lane M: <t>DNA</t> marker DL 2000, lane 1: PCR amplification of cbpA in Δ cbpA mutants, lane 2: PCR amplification of cbpA in wild type strain, lane 3: PCR amplification of nam - amidase in wild type strain, lane 4: PCR amplification of nam - amidase in Δ nam - amidase mutants. ( E ) Adhesion of L. salivarius REN, Δ cbpA and Δ nam - amidase mutants to the HT-29 cells. Data were analyzed by the paired Student’s t test (* P
    Fastking Gdna Dispelling Rt Supermix, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 89/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co biology tissue gdna extraction kit
    Identification of S-layer proteins of L. <t>salivarius</t> REN. ( A ) SDS-PAGE of cell surface proteins extracted from L. salivarius REN. ( B ) Gene organization of the region surrounding the cbpA and nam - amidase locus. Arrows indicate the locations of primers used for PCR analysis. The red shaded regions represented the deletion target. The green and blue shaded regions represented the upstream and downstream fragments in target genes. The chromosomal map is not drawn to scale. ( C ) Schematic overview of the construction of Δ cbpA and Δ nam - amidase mutants. ( D ) PCR identification of double crossover recombinant. Lane M: <t>DNA</t> marker DL 2000, lane 1: PCR amplification of cbpA in Δ cbpA mutants, lane 2: PCR amplification of cbpA in wild type strain, lane 3: PCR amplification of nam - amidase in wild type strain, lane 4: PCR amplification of nam - amidase in Δ nam - amidase mutants. ( E ) Adhesion of L. salivarius REN, Δ cbpA and Δ nam - amidase mutants to the HT-29 cells. Data were analyzed by the paired Student’s t test (* P
    Biology Tissue Gdna Extraction Kit, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 86/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co hifi script quick gdna removal kit
    Identification of S-layer proteins of L. <t>salivarius</t> REN. ( A ) SDS-PAGE of cell surface proteins extracted from L. salivarius REN. ( B ) Gene organization of the region surrounding the cbpA and nam - amidase locus. Arrows indicate the locations of primers used for PCR analysis. The red shaded regions represented the deletion target. The green and blue shaded regions represented the upstream and downstream fragments in target genes. The chromosomal map is not drawn to scale. ( C ) Schematic overview of the construction of Δ cbpA and Δ nam - amidase mutants. ( D ) PCR identification of double crossover recombinant. Lane M: <t>DNA</t> marker DL 2000, lane 1: PCR amplification of cbpA in Δ cbpA mutants, lane 2: PCR amplification of cbpA in wild type strain, lane 3: PCR amplification of nam - amidase in wild type strain, lane 4: PCR amplification of nam - amidase in Δ nam - amidase mutants. ( E ) Adhesion of L. salivarius REN, Δ cbpA and Δ nam - amidase mutants to the HT-29 cells. Data were analyzed by the paired Student’s t test (* P
    Hifi Script Quick Gdna Removal Kit, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 80/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co fastking gdna dispelling rt supermix kit
    Identification of S-layer proteins of L. <t>salivarius</t> REN. ( A ) SDS-PAGE of cell surface proteins extracted from L. salivarius REN. ( B ) Gene organization of the region surrounding the cbpA and nam - amidase locus. Arrows indicate the locations of primers used for PCR analysis. The red shaded regions represented the deletion target. The green and blue shaded regions represented the upstream and downstream fragments in target genes. The chromosomal map is not drawn to scale. ( C ) Schematic overview of the construction of Δ cbpA and Δ nam - amidase mutants. ( D ) PCR identification of double crossover recombinant. Lane M: <t>DNA</t> marker DL 2000, lane 1: PCR amplification of cbpA in Δ cbpA mutants, lane 2: PCR amplification of cbpA in wild type strain, lane 3: PCR amplification of nam - amidase in wild type strain, lane 4: PCR amplification of nam - amidase in Δ nam - amidase mutants. ( E ) Adhesion of L. salivarius REN, Δ cbpA and Δ nam - amidase mutants to the HT-29 cells. Data were analyzed by the paired Student’s t test (* P
    Fastking Gdna Dispelling Rt Supermix Kit, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 84/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co genomic dna kit
    Vaccination with CpG adjuvant reduces <t>HBV</t> <t>DNA</t> in the liver. ( a ) AAV/HBV (2×10 10 vg)-infected mice were treated with the indicated vaccine combinations. HBV DNA levels in the liver were determined by real-time PCR at 11 weeks post-vaccination. ( b ) HBsAg levels in the liver were measured by ELISA at 11 weeks post-vaccination with homogenized liver tissues. ( c ) Immunohistochemical staining of HBcAg in the liver sections of different groups ( n =3). AAV, adeno-associated virus; HBcAg, HBV core antigen; HBsAg, HBV surface antigen; HBV, hepatitis B virus.
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    tiangen biotech co tianamp genomic dna kit
    Vaccination with CpG adjuvant reduces <t>HBV</t> <t>DNA</t> in the liver. ( a ) AAV/HBV (2×10 10 vg)-infected mice were treated with the indicated vaccine combinations. HBV DNA levels in the liver were determined by real-time PCR at 11 weeks post-vaccination. ( b ) HBsAg levels in the liver were measured by ELISA at 11 weeks post-vaccination with homogenized liver tissues. ( c ) Immunohistochemical staining of HBcAg in the liver sections of different groups ( n =3). AAV, adeno-associated virus; HBcAg, HBV core antigen; HBsAg, HBV surface antigen; HBV, hepatitis B virus.
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    tiangen biotech co genomic dna genomic dna
    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
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    tiangen biotech co gdna dispelling rt supermix rt reagent kit
    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
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    tiangen biotech co genomic dna isolation genomic dna
    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
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    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
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    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
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    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
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    tiangen biotech co tianamp bacterial genomic dna kit
    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
    Tianamp Bacterial Genomic Dna Kit, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 86/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
    Bacterial Genomic Dna Purification Kit, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 77/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
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    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
    Tguide Blood Genomic Dna Kit, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
    Tianamp Genomic Dna Purification Kit, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 85/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co genomic dna extraction bacterial genomic dna
    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
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    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
    Genomic Dna Plant Genomic Dna, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 77/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effects of <t>Lfcin4</t> and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine <t>(DNA)</t> (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).
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    TALENs-mediated homozygous recombination. (A) T7E1 enzyme analysis of transfected CD4 + U87 cells after each <t>transfection</t> with CCR5-TALENs. The targeting efficiencies after first, second and third rounds of transfection were 14.80, 38.20 and 50.04%, respectively. M, 100 bp marker; wild-type, wild-type cells; lanes 1–3, transfected cells after the first, second and third rounds of transfections. (B) PCR of CCR5Δ32/Δ1 genotype cells. M, 100 bp marker; lane 1, prior to transfection; lane 2, after CCR5-TALENs transfection; lane 3, after CCR5Δ1-TALENs transfection; the CCR5Δ32 band (168 bp) was 2.87-times brighter than the CCR5Δ1 band; lane 4, wild-type cells (negative control); lane 5, CCR5Δ32 donor <t>DNA</t> (positive control). (C) Sequencing of wild-type CD4 + U87 cells and those with CCR5Δ32/Δ1 mutations. (D) PCR of 34 single-cell cultured clones of CD4 + U87 cells with CCR5Δ32/Δ1 mutations post-transfection with CCR5Δ1-TALENs. The CCR5Δ32 band was observed in all clones; however, as a single band, it was only seen in clones 16, 18 and 25 post-transfection with CCR5Δ1-TALENs. M, 100 bp marker; lanes 1–34, single-cell cultured cells post-transfection with CCR5Δ1-TALENs; wild-type, wild-type cells (negative control); donor, CCR5Δ32 donor DNA (positive control). (E) Gene sequencing of clones 16, 18 and 25. CCR5, C-C chemokine receptor type 5; CD4, cluster of differentiation 4; PCR, polymerase chain reaction; T7E1, T7 endonuclease 1; TALENs, transcription activator-like effector nucleases.
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    TALENs-mediated homozygous recombination. (A) T7E1 enzyme analysis of transfected CD4 + U87 cells after each <t>transfection</t> with CCR5-TALENs. The targeting efficiencies after first, second and third rounds of transfection were 14.80, 38.20 and 50.04%, respectively. M, 100 bp marker; wild-type, wild-type cells; lanes 1–3, transfected cells after the first, second and third rounds of transfections. (B) PCR of CCR5Δ32/Δ1 genotype cells. M, 100 bp marker; lane 1, prior to transfection; lane 2, after CCR5-TALENs transfection; lane 3, after CCR5Δ1-TALENs transfection; the CCR5Δ32 band (168 bp) was 2.87-times brighter than the CCR5Δ1 band; lane 4, wild-type cells (negative control); lane 5, CCR5Δ32 donor <t>DNA</t> (positive control). (C) Sequencing of wild-type CD4 + U87 cells and those with CCR5Δ32/Δ1 mutations. (D) PCR of 34 single-cell cultured clones of CD4 + U87 cells with CCR5Δ32/Δ1 mutations post-transfection with CCR5Δ1-TALENs. The CCR5Δ32 band was observed in all clones; however, as a single band, it was only seen in clones 16, 18 and 25 post-transfection with CCR5Δ1-TALENs. M, 100 bp marker; lanes 1–34, single-cell cultured cells post-transfection with CCR5Δ1-TALENs; wild-type, wild-type cells (negative control); donor, CCR5Δ32 donor DNA (positive control). (E) Gene sequencing of clones 16, 18 and 25. CCR5, C-C chemokine receptor type 5; CD4, cluster of differentiation 4; PCR, polymerase chain reaction; T7E1, T7 endonuclease 1; TALENs, transcription activator-like effector nucleases.
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    TALENs-mediated homozygous recombination. (A) T7E1 enzyme analysis of transfected CD4 + U87 cells after each <t>transfection</t> with CCR5-TALENs. The targeting efficiencies after first, second and third rounds of transfection were 14.80, 38.20 and 50.04%, respectively. M, 100 bp marker; wild-type, wild-type cells; lanes 1–3, transfected cells after the first, second and third rounds of transfections. (B) PCR of CCR5Δ32/Δ1 genotype cells. M, 100 bp marker; lane 1, prior to transfection; lane 2, after CCR5-TALENs transfection; lane 3, after CCR5Δ1-TALENs transfection; the CCR5Δ32 band (168 bp) was 2.87-times brighter than the CCR5Δ1 band; lane 4, wild-type cells (negative control); lane 5, CCR5Δ32 donor <t>DNA</t> (positive control). (C) Sequencing of wild-type CD4 + U87 cells and those with CCR5Δ32/Δ1 mutations. (D) PCR of 34 single-cell cultured clones of CD4 + U87 cells with CCR5Δ32/Δ1 mutations post-transfection with CCR5Δ1-TALENs. The CCR5Δ32 band was observed in all clones; however, as a single band, it was only seen in clones 16, 18 and 25 post-transfection with CCR5Δ1-TALENs. M, 100 bp marker; lanes 1–34, single-cell cultured cells post-transfection with CCR5Δ1-TALENs; wild-type, wild-type cells (negative control); donor, CCR5Δ32 donor DNA (positive control). (E) Gene sequencing of clones 16, 18 and 25. CCR5, C-C chemokine receptor type 5; CD4, cluster of differentiation 4; PCR, polymerase chain reaction; T7E1, T7 endonuclease 1; TALENs, transcription activator-like effector nucleases.
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    TALENs-mediated homozygous recombination. (A) T7E1 enzyme analysis of transfected CD4 + U87 cells after each <t>transfection</t> with CCR5-TALENs. The targeting efficiencies after first, second and third rounds of transfection were 14.80, 38.20 and 50.04%, respectively. M, 100 bp marker; wild-type, wild-type cells; lanes 1–3, transfected cells after the first, second and third rounds of transfections. (B) PCR of CCR5Δ32/Δ1 genotype cells. M, 100 bp marker; lane 1, prior to transfection; lane 2, after CCR5-TALENs transfection; lane 3, after CCR5Δ1-TALENs transfection; the CCR5Δ32 band (168 bp) was 2.87-times brighter than the CCR5Δ1 band; lane 4, wild-type cells (negative control); lane 5, CCR5Δ32 donor <t>DNA</t> (positive control). (C) Sequencing of wild-type CD4 + U87 cells and those with CCR5Δ32/Δ1 mutations. (D) PCR of 34 single-cell cultured clones of CD4 + U87 cells with CCR5Δ32/Δ1 mutations post-transfection with CCR5Δ1-TALENs. The CCR5Δ32 band was observed in all clones; however, as a single band, it was only seen in clones 16, 18 and 25 post-transfection with CCR5Δ1-TALENs. M, 100 bp marker; lanes 1–34, single-cell cultured cells post-transfection with CCR5Δ1-TALENs; wild-type, wild-type cells (negative control); donor, CCR5Δ32 donor DNA (positive control). (E) Gene sequencing of clones 16, 18 and 25. CCR5, C-C chemokine receptor type 5; CD4, cluster of differentiation 4; PCR, polymerase chain reaction; T7E1, T7 endonuclease 1; TALENs, transcription activator-like effector nucleases.
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    TALENs-mediated homozygous recombination. (A) T7E1 enzyme analysis of transfected CD4 + U87 cells after each <t>transfection</t> with CCR5-TALENs. The targeting efficiencies after first, second and third rounds of transfection were 14.80, 38.20 and 50.04%, respectively. M, 100 bp marker; wild-type, wild-type cells; lanes 1–3, transfected cells after the first, second and third rounds of transfections. (B) PCR of CCR5Δ32/Δ1 genotype cells. M, 100 bp marker; lane 1, prior to transfection; lane 2, after CCR5-TALENs transfection; lane 3, after CCR5Δ1-TALENs transfection; the CCR5Δ32 band (168 bp) was 2.87-times brighter than the CCR5Δ1 band; lane 4, wild-type cells (negative control); lane 5, CCR5Δ32 donor <t>DNA</t> (positive control). (C) Sequencing of wild-type CD4 + U87 cells and those with CCR5Δ32/Δ1 mutations. (D) PCR of 34 single-cell cultured clones of CD4 + U87 cells with CCR5Δ32/Δ1 mutations post-transfection with CCR5Δ1-TALENs. The CCR5Δ32 band was observed in all clones; however, as a single band, it was only seen in clones 16, 18 and 25 post-transfection with CCR5Δ1-TALENs. M, 100 bp marker; lanes 1–34, single-cell cultured cells post-transfection with CCR5Δ1-TALENs; wild-type, wild-type cells (negative control); donor, CCR5Δ32 donor DNA (positive control). (E) Gene sequencing of clones 16, 18 and 25. CCR5, C-C chemokine receptor type 5; CD4, cluster of differentiation 4; PCR, polymerase chain reaction; T7E1, T7 endonuclease 1; TALENs, transcription activator-like effector nucleases.
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    TALENs-mediated homozygous recombination. (A) T7E1 enzyme analysis of transfected CD4 + U87 cells after each <t>transfection</t> with CCR5-TALENs. The targeting efficiencies after first, second and third rounds of transfection were 14.80, 38.20 and 50.04%, respectively. M, 100 bp marker; wild-type, wild-type cells; lanes 1–3, transfected cells after the first, second and third rounds of transfections. (B) PCR of CCR5Δ32/Δ1 genotype cells. M, 100 bp marker; lane 1, prior to transfection; lane 2, after CCR5-TALENs transfection; lane 3, after CCR5Δ1-TALENs transfection; the CCR5Δ32 band (168 bp) was 2.87-times brighter than the CCR5Δ1 band; lane 4, wild-type cells (negative control); lane 5, CCR5Δ32 donor <t>DNA</t> (positive control). (C) Sequencing of wild-type CD4 + U87 cells and those with CCR5Δ32/Δ1 mutations. (D) PCR of 34 single-cell cultured clones of CD4 + U87 cells with CCR5Δ32/Δ1 mutations post-transfection with CCR5Δ1-TALENs. The CCR5Δ32 band was observed in all clones; however, as a single band, it was only seen in clones 16, 18 and 25 post-transfection with CCR5Δ1-TALENs. M, 100 bp marker; lanes 1–34, single-cell cultured cells post-transfection with CCR5Δ1-TALENs; wild-type, wild-type cells (negative control); donor, CCR5Δ32 donor DNA (positive control). (E) Gene sequencing of clones 16, 18 and 25. CCR5, C-C chemokine receptor type 5; CD4, cluster of differentiation 4; PCR, polymerase chain reaction; T7E1, T7 endonuclease 1; TALENs, transcription activator-like effector nucleases.
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    TALENs-mediated homozygous recombination. (A) T7E1 enzyme analysis of transfected CD4 + U87 cells after each <t>transfection</t> with CCR5-TALENs. The targeting efficiencies after first, second and third rounds of transfection were 14.80, 38.20 and 50.04%, respectively. M, 100 bp marker; wild-type, wild-type cells; lanes 1–3, transfected cells after the first, second and third rounds of transfections. (B) PCR of CCR5Δ32/Δ1 genotype cells. M, 100 bp marker; lane 1, prior to transfection; lane 2, after CCR5-TALENs transfection; lane 3, after CCR5Δ1-TALENs transfection; the CCR5Δ32 band (168 bp) was 2.87-times brighter than the CCR5Δ1 band; lane 4, wild-type cells (negative control); lane 5, CCR5Δ32 donor <t>DNA</t> (positive control). (C) Sequencing of wild-type CD4 + U87 cells and those with CCR5Δ32/Δ1 mutations. (D) PCR of 34 single-cell cultured clones of CD4 + U87 cells with CCR5Δ32/Δ1 mutations post-transfection with CCR5Δ1-TALENs. The CCR5Δ32 band was observed in all clones; however, as a single band, it was only seen in clones 16, 18 and 25 post-transfection with CCR5Δ1-TALENs. M, 100 bp marker; lanes 1–34, single-cell cultured cells post-transfection with CCR5Δ1-TALENs; wild-type, wild-type cells (negative control); donor, CCR5Δ32 donor DNA (positive control). (E) Gene sequencing of clones 16, 18 and 25. CCR5, C-C chemokine receptor type 5; CD4, cluster of differentiation 4; PCR, polymerase chain reaction; T7E1, T7 endonuclease 1; TALENs, transcription activator-like effector nucleases.
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    DNA identification and gene transcripts analysis . A) PCR analyses of transgenic hairy root lines. Analyses for the presence of AaPMT (1269 bp) and AaTRI genes (1074 bp) in PT lines (1), AaPMT gene in P lines (2) and AaTRI gene in T lines (3), respectively. M, DL-2000 Marker. PC, positive control (vector contains corresponding gene). BC, blank control (hairy root generate from blank-vector transformation). B) Effects of transformation on the expression of related genes. AaPMT T and AaTRI T represent total gene of AaPMT and AaTRI respectively, AaPMT E and AaTRI E represent endogenous AaPMT and AaTRI respectively. C) Real-time fluorescence quantitative PCR analysis of the expressions of AaPMT and AaTRI in transgenic hairy roots.

    Journal: BMC Biotechnology

    Article Title: Co-expression of AaPMT and AaTRI effectively enhances the yields of tropane alkaloids in Anisodus acutangulus hairy roots

    doi: 10.1186/1472-6750-11-43

    Figure Lengend Snippet: DNA identification and gene transcripts analysis . A) PCR analyses of transgenic hairy root lines. Analyses for the presence of AaPMT (1269 bp) and AaTRI genes (1074 bp) in PT lines (1), AaPMT gene in P lines (2) and AaTRI gene in T lines (3), respectively. M, DL-2000 Marker. PC, positive control (vector contains corresponding gene). BC, blank control (hairy root generate from blank-vector transformation). B) Effects of transformation on the expression of related genes. AaPMT T and AaTRI T represent total gene of AaPMT and AaTRI respectively, AaPMT E and AaTRI E represent endogenous AaPMT and AaTRI respectively. C) Real-time fluorescence quantitative PCR analysis of the expressions of AaPMT and AaTRI in transgenic hairy roots.

    Article Snippet: DNA extraction and PCR analysis When the engineered hairy roots grew to about 5 cm, genomic DNA was isolated from hairy root samples by using DNA pure Plant Kit (Tiangen Biotech Co., Ltd, Beijing, China), which was used in PCR analysis for detecting the presence of AaPMT and/or AaTRI in transgenic hairy root cultures (Table ).

    Techniques: Polymerase Chain Reaction, Transgenic Assay, Marker, Positive Control, Plasmid Preparation, Transformation Assay, Expressing, Fluorescence, Real-time Polymerase Chain Reaction

    Characterization of B. amyloliquefaciens strain HAB-2 and its mutant. a Gel electrophoresis of PCR products of wild-type strain HAB-2 (lane HAB-2) and its mutant (lane Mutant); b MALDI-TOF MS analysis on the extract of HAB△ lpa mutant, 3000 bp DNA marker

    Journal: BMC Microbiology

    Article Title: Characterization of lpaH2 gene corresponding to lipopeptide synthesis in Bacillus amyloliquefaciens HAB-2

    doi: 10.1186/s12866-017-1134-z

    Figure Lengend Snippet: Characterization of B. amyloliquefaciens strain HAB-2 and its mutant. a Gel electrophoresis of PCR products of wild-type strain HAB-2 (lane HAB-2) and its mutant (lane Mutant); b MALDI-TOF MS analysis on the extract of HAB△ lpa mutant, 3000 bp DNA marker

    Article Snippet: The chromosomal DNA of strain HAB-2 was extracted using Bacterial Genomic DNA Extraction Kit (Tiangen Biochemical Science and Technology Co., LTD) according to the manufacturer’s protocol.

    Techniques: Mutagenesis, Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Mass Spectrometry, Marker

    Lipopeptide analysis from B. amyloliquefaciens HAB-2 and B. subtilis 168. a Amplification of lpa and sfp gene fragments from B. amyloliquefaciens HAB-2 and B. subtilis 168. From lane 1 to 4: HAB-2- sfp ; B. subtilis 168- sfp ; HAB-2- lpa ; and B. subtilis 168- lpa . Lane M: 2000 bp DNA marker. b Southern-blot hybridization for lpaH2 and sfp genes, a: lpa from the strain HAB-2, b: chromosomal DNA of the strain HAB-2, c: chromosomal DNA of B. subtilis 168, d: sfp from B. subtilis 168, e: chromosomal DNA of B. subtilis 168, f: chromosomal DNA of the strain HAB-2. c Antifungal activity of B. amyloliquefaciens HAB-2 and B. subtilis 168 against C. gloeosporioides on agar plate. d Inhibitory effect of lipopetides produced from B. amyloliquefaciens HAB-2 against C. gloeosporioides

    Journal: BMC Microbiology

    Article Title: Characterization of lpaH2 gene corresponding to lipopeptide synthesis in Bacillus amyloliquefaciens HAB-2

    doi: 10.1186/s12866-017-1134-z

    Figure Lengend Snippet: Lipopeptide analysis from B. amyloliquefaciens HAB-2 and B. subtilis 168. a Amplification of lpa and sfp gene fragments from B. amyloliquefaciens HAB-2 and B. subtilis 168. From lane 1 to 4: HAB-2- sfp ; B. subtilis 168- sfp ; HAB-2- lpa ; and B. subtilis 168- lpa . Lane M: 2000 bp DNA marker. b Southern-blot hybridization for lpaH2 and sfp genes, a: lpa from the strain HAB-2, b: chromosomal DNA of the strain HAB-2, c: chromosomal DNA of B. subtilis 168, d: sfp from B. subtilis 168, e: chromosomal DNA of B. subtilis 168, f: chromosomal DNA of the strain HAB-2. c Antifungal activity of B. amyloliquefaciens HAB-2 and B. subtilis 168 against C. gloeosporioides on agar plate. d Inhibitory effect of lipopetides produced from B. amyloliquefaciens HAB-2 against C. gloeosporioides

    Article Snippet: The chromosomal DNA of strain HAB-2 was extracted using Bacterial Genomic DNA Extraction Kit (Tiangen Biochemical Science and Technology Co., LTD) according to the manufacturer’s protocol.

    Techniques: Amplification, Marker, Southern Blot, Hybridization, Activity Assay, Produced

    High-throughput sequencing analysis of CRISPR/Cas9-Induced mutagenesis in three BnSPL3 homoeologs (A–C) , statistical analysis of editing frequency occurred in rapeseed SPL3A, SPL3B , and SPL3C genomic sites, respectively. Rapeseed sample 1#, 2#, 20#, 24#, 41# were from independent transgenic events. Excepting transgenic line 24#, all the rest samples display obvious heteroduplexed DNA bands in PAGE-based assays. (D–F) Sequences alignment of different mutation types identified from high-throughput sequencing analysis.

    Journal: Frontiers in Plant Science

    Article Title: An Efficient CRISPR/Cas9 Platform for Rapidly Generating Simultaneous Mutagenesis of Multiple Gene Homoeologs in Allotetraploid Oilseed Rape

    doi: 10.3389/fpls.2018.00442

    Figure Lengend Snippet: High-throughput sequencing analysis of CRISPR/Cas9-Induced mutagenesis in three BnSPL3 homoeologs (A–C) , statistical analysis of editing frequency occurred in rapeseed SPL3A, SPL3B , and SPL3C genomic sites, respectively. Rapeseed sample 1#, 2#, 20#, 24#, 41# were from independent transgenic events. Excepting transgenic line 24#, all the rest samples display obvious heteroduplexed DNA bands in PAGE-based assays. (D–F) Sequences alignment of different mutation types identified from high-throughput sequencing analysis.

    Article Snippet: Genomic DNA was extracted from transgenic rapeseed plants using the Plant Genomic DNA Kit (TIANGEN, China).

    Techniques: Next-Generation Sequencing, CRISPR, Mutagenesis, Transgenic Assay, Polyacrylamide Gel Electrophoresis

    Detection of BRAF gene mutation at exons 11 and 15. (A) Gel electrophoresis of the PCR products of exon 11 and 15 of the BRAF gene. The PCR product size of exon 11 is 356 bp, and that of exon 15 is 249 bp. Part of a sequence chromatogram from the Sanger sequencing of BRAF from (B) a patient and (C) the positive control. The red arrow indicates a BRAF mutation (T1799A) of exon 15 in the BCPAP papillary thyroid cancer cell line. BRAF, B-Raf proto-oncogene, serine/threonine kinase; PCR, polymerase chain reaction; M, DNA marker; NE, normal endometrium; EU, eutopic endometrium of endometriosis; EC, ectopic endometrium of endometriosis.

    Journal: International Journal of Molecular Medicine

    Article Title: Analysis of the oncogene BRAF mutation and the correlation of the expression of wild-type BRAF and CREB1 in endometriosis

    doi: 10.3892/ijmm.2017.3342

    Figure Lengend Snippet: Detection of BRAF gene mutation at exons 11 and 15. (A) Gel electrophoresis of the PCR products of exon 11 and 15 of the BRAF gene. The PCR product size of exon 11 is 356 bp, and that of exon 15 is 249 bp. Part of a sequence chromatogram from the Sanger sequencing of BRAF from (B) a patient and (C) the positive control. The red arrow indicates a BRAF mutation (T1799A) of exon 15 in the BCPAP papillary thyroid cancer cell line. BRAF, B-Raf proto-oncogene, serine/threonine kinase; PCR, polymerase chain reaction; M, DNA marker; NE, normal endometrium; EU, eutopic endometrium of endometriosis; EC, ectopic endometrium of endometriosis.

    Article Snippet: Genomic DNA isolation, PCR and direct sequencing Genomic DNA was extracted from freshly frozen tissue (100 mg) using the TIANamp Genomic DNA kit (Tiangen Biotech Co., Ltd., Beijing, China) according to the manufacturer's protocol.

    Techniques: Mutagenesis, Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Sequencing, Positive Control, Marker

    PCR detection of 22 putative transgenic plants. 1~ 22 are putative transgenic plants, positive control (plasmid DNA as template, CK+) and negative control (non-transgenic plants, CK-)

    Journal: Hereditas

    Article Title: Enhancement of grain number per spike by RNA interference of cytokinin oxidase 2 gene in bread wheat

    doi: 10.1186/s41065-018-0071-7

    Figure Lengend Snippet: PCR detection of 22 putative transgenic plants. 1~ 22 are putative transgenic plants, positive control (plasmid DNA as template, CK+) and negative control (non-transgenic plants, CK-)

    Article Snippet: Genomic DNA was extracted from young leaf tissues of receptor NB1 and transgenic plants using plant genomic DNA Extraction Kit (Tiangen Biotech, Beijing, China).

    Techniques: Polymerase Chain Reaction, Transgenic Assay, Positive Control, Plasmid Preparation, Negative Control

    Southern blotting analysis in selected T 0 primary wheat transformants. Genomic DNA digested with Eco R V and hybridized with FAD2 probe. The number of reactive bands in each lane represents the transgene copies in each transgenic line. Lane 1 is WT (NB1), lanes 2~ 6 are JW1-1A, JW5-1A, JW41-1B, JW39-3A and JW1-2B, respectively, M is the marker: λ-EcoT14 I digest (TaKaRa, Dalian, China)

    Journal: Hereditas

    Article Title: Enhancement of grain number per spike by RNA interference of cytokinin oxidase 2 gene in bread wheat

    doi: 10.1186/s41065-018-0071-7

    Figure Lengend Snippet: Southern blotting analysis in selected T 0 primary wheat transformants. Genomic DNA digested with Eco R V and hybridized with FAD2 probe. The number of reactive bands in each lane represents the transgene copies in each transgenic line. Lane 1 is WT (NB1), lanes 2~ 6 are JW1-1A, JW5-1A, JW41-1B, JW39-3A and JW1-2B, respectively, M is the marker: λ-EcoT14 I digest (TaKaRa, Dalian, China)

    Article Snippet: Genomic DNA was extracted from young leaf tissues of receptor NB1 and transgenic plants using plant genomic DNA Extraction Kit (Tiangen Biotech, Beijing, China).

    Techniques: Southern Blot, Transgenic Assay, Marker

    Representative variation of DNA methylation pattern. H ( Eco R I + Hpa II digest) and M ( Eco R I + Msp I digest) refer to digestion with Eco R I + Hpa II and Eco R I + Msp I, respectively. “→” red arrows represent parts of differential methylated bands between common wheat and wheat carrying Gc chromosome(s). M stands for Marker DL2000. MA (1, 1), presence in both H ( Eco R I + Hpa II digest) and M ( Eco R I + Msp I digest) lanes; MB (1, 0), presence in H and absence in M lane; MC (0, 1), absence in H but presence in M lane; MD (0, 0) absence in both H and M lanes. CS: T. aestivum cv. Chinese Spring. CS–3C: monosomic addition line of Chinese Spring (CS) that carries a gametocidal chromosome 3C originated from Aegilops triuncialis . CS–3C3C: disomic addition line of Chinese Spring (CS) that carries two gametocidal chromosome 3C originated from Aegilops triuncialis .

    Journal: International Journal of Molecular Sciences

    Article Title: The Variation Analysis of DNA Methylation in Wheat Carrying Gametocidal Chromosome 3C from Aegilops triuncialis

    doi: 10.3390/ijms18081738

    Figure Lengend Snippet: Representative variation of DNA methylation pattern. H ( Eco R I + Hpa II digest) and M ( Eco R I + Msp I digest) refer to digestion with Eco R I + Hpa II and Eco R I + Msp I, respectively. “→” red arrows represent parts of differential methylated bands between common wheat and wheat carrying Gc chromosome(s). M stands for Marker DL2000. MA (1, 1), presence in both H ( Eco R I + Hpa II digest) and M ( Eco R I + Msp I digest) lanes; MB (1, 0), presence in H and absence in M lane; MC (0, 1), absence in H but presence in M lane; MD (0, 0) absence in both H and M lanes. CS: T. aestivum cv. Chinese Spring. CS–3C: monosomic addition line of Chinese Spring (CS) that carries a gametocidal chromosome 3C originated from Aegilops triuncialis . CS–3C3C: disomic addition line of Chinese Spring (CS) that carries two gametocidal chromosome 3C originated from Aegilops triuncialis .

    Article Snippet: DNA Extraction Genomic DNA was extracted from anthers of CS, CS–3C and CS–3C3C using DNAsecure Plant Kit (TIANGEN, DP320, Beijing, China) according to the manufacturer’s instructions.

    Techniques: DNA Methylation Assay, Methylation, Marker

    Identification of S-layer proteins of L. salivarius REN. ( A ) SDS-PAGE of cell surface proteins extracted from L. salivarius REN. ( B ) Gene organization of the region surrounding the cbpA and nam - amidase locus. Arrows indicate the locations of primers used for PCR analysis. The red shaded regions represented the deletion target. The green and blue shaded regions represented the upstream and downstream fragments in target genes. The chromosomal map is not drawn to scale. ( C ) Schematic overview of the construction of Δ cbpA and Δ nam - amidase mutants. ( D ) PCR identification of double crossover recombinant. Lane M: DNA marker DL 2000, lane 1: PCR amplification of cbpA in Δ cbpA mutants, lane 2: PCR amplification of cbpA in wild type strain, lane 3: PCR amplification of nam - amidase in wild type strain, lane 4: PCR amplification of nam - amidase in Δ nam - amidase mutants. ( E ) Adhesion of L. salivarius REN, Δ cbpA and Δ nam - amidase mutants to the HT-29 cells. Data were analyzed by the paired Student’s t test (* P

    Journal: Scientific Reports

    Article Title: The Adhesion of Lactobacillus salivarius REN to a Human Intestinal Epithelial Cell Line Requires S-layer Proteins

    doi: 10.1038/srep44029

    Figure Lengend Snippet: Identification of S-layer proteins of L. salivarius REN. ( A ) SDS-PAGE of cell surface proteins extracted from L. salivarius REN. ( B ) Gene organization of the region surrounding the cbpA and nam - amidase locus. Arrows indicate the locations of primers used for PCR analysis. The red shaded regions represented the deletion target. The green and blue shaded regions represented the upstream and downstream fragments in target genes. The chromosomal map is not drawn to scale. ( C ) Schematic overview of the construction of Δ cbpA and Δ nam - amidase mutants. ( D ) PCR identification of double crossover recombinant. Lane M: DNA marker DL 2000, lane 1: PCR amplification of cbpA in Δ cbpA mutants, lane 2: PCR amplification of cbpA in wild type strain, lane 3: PCR amplification of nam - amidase in wild type strain, lane 4: PCR amplification of nam - amidase in Δ nam - amidase mutants. ( E ) Adhesion of L. salivarius REN, Δ cbpA and Δ nam - amidase mutants to the HT-29 cells. Data were analyzed by the paired Student’s t test (* P

    Article Snippet: Genomic DNA of L. salivarius REN was isolated using a genomic DNA purification kit (Tiangen Biotech Co., Ltd., Beijing, China).

    Techniques: SDS Page, Polymerase Chain Reaction, Recombinant, Marker, Amplification

    Vaccination with CpG adjuvant reduces HBV DNA in the liver. ( a ) AAV/HBV (2×10 10 vg)-infected mice were treated with the indicated vaccine combinations. HBV DNA levels in the liver were determined by real-time PCR at 11 weeks post-vaccination. ( b ) HBsAg levels in the liver were measured by ELISA at 11 weeks post-vaccination with homogenized liver tissues. ( c ) Immunohistochemical staining of HBcAg in the liver sections of different groups ( n =3). AAV, adeno-associated virus; HBcAg, HBV core antigen; HBsAg, HBV surface antigen; HBV, hepatitis B virus.

    Journal: Cellular and Molecular Immunology

    Article Title: A mouse model for HBV immunotolerance and immunotherapy

    doi: 10.1038/cmi.2013.43

    Figure Lengend Snippet: Vaccination with CpG adjuvant reduces HBV DNA in the liver. ( a ) AAV/HBV (2×10 10 vg)-infected mice were treated with the indicated vaccine combinations. HBV DNA levels in the liver were determined by real-time PCR at 11 weeks post-vaccination. ( b ) HBsAg levels in the liver were measured by ELISA at 11 weeks post-vaccination with homogenized liver tissues. ( c ) Immunohistochemical staining of HBcAg in the liver sections of different groups ( n =3). AAV, adeno-associated virus; HBcAg, HBV core antigen; HBsAg, HBV surface antigen; HBV, hepatitis B virus.

    Article Snippet: Liver HBV DNA was extracted from 50 mg of liver tissues using a genomic DNA kit (TIANGEN Biotech, Beijing, China).

    Techniques: Infection, Mouse Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining

    AAV/HBV inoculation resulted in persistent HBV viremia in immunocompetent mice. ( a ) Female or male C57BL/6 mice ( n =6, 6–8 weeks old) were infected with AAV/HBV at 5×10 10 viral genome equivalents (vg) through tail vein injection. At 14 days post-infection, blood samples were collected and serum HBsAg was measured by ELISA. ( b ) Intraliver injection of 3-day-old neonatal C57BL/6 mice ( n =6–7) was performed with 1×10 10 or 2×10 10 vg AAV/HBV, and serum HBsAg titers were examined at 48 days post-infection. ( c ) C57BL/6 mice ( n =6, 6–8 weeks old) were infected with AAV/HBV at 2×10 10 , 5×10 10 or 1×10 11 vg. The mice were bled biweekly after infection. Serum HBsAg was measured by ELISA. The lower limit of detection was 0.5 ng/ml. ( d ) Adult C57BL/6 mice were intravenously injected with 2×10 10 , 5×10 10 or 1×10 11 vg virus and were bled biweekly to monitor serum HBeAg titers. ( e ) On day 40 post-AAV/HBV infection, HBV genomic DNA in the serum was determined by real-time PCR. ( f ) Immunohistochemical staining of HBcAg in the liver of AAV/HBV infected mice (12 weeks post-infection). AAV, adeno-associated virus; HBcAg, HBV core antigen; HBsAg, HBV surface antigen; HBV, hepatitis B virus.

    Journal: Cellular and Molecular Immunology

    Article Title: A mouse model for HBV immunotolerance and immunotherapy

    doi: 10.1038/cmi.2013.43

    Figure Lengend Snippet: AAV/HBV inoculation resulted in persistent HBV viremia in immunocompetent mice. ( a ) Female or male C57BL/6 mice ( n =6, 6–8 weeks old) were infected with AAV/HBV at 5×10 10 viral genome equivalents (vg) through tail vein injection. At 14 days post-infection, blood samples were collected and serum HBsAg was measured by ELISA. ( b ) Intraliver injection of 3-day-old neonatal C57BL/6 mice ( n =6–7) was performed with 1×10 10 or 2×10 10 vg AAV/HBV, and serum HBsAg titers were examined at 48 days post-infection. ( c ) C57BL/6 mice ( n =6, 6–8 weeks old) were infected with AAV/HBV at 2×10 10 , 5×10 10 or 1×10 11 vg. The mice were bled biweekly after infection. Serum HBsAg was measured by ELISA. The lower limit of detection was 0.5 ng/ml. ( d ) Adult C57BL/6 mice were intravenously injected with 2×10 10 , 5×10 10 or 1×10 11 vg virus and were bled biweekly to monitor serum HBeAg titers. ( e ) On day 40 post-AAV/HBV infection, HBV genomic DNA in the serum was determined by real-time PCR. ( f ) Immunohistochemical staining of HBcAg in the liver of AAV/HBV infected mice (12 weeks post-infection). AAV, adeno-associated virus; HBcAg, HBV core antigen; HBsAg, HBV surface antigen; HBV, hepatitis B virus.

    Article Snippet: Liver HBV DNA was extracted from 50 mg of liver tissues using a genomic DNA kit (TIANGEN Biotech, Beijing, China).

    Techniques: Mouse Assay, Infection, Injection, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Immunohistochemistry, Staining

    The immune response was undetectable in AAV/HBV-infected mice. ( a ) Serum HBsAb levels in C57BL/6 mice receiving 15 µg of pBlue-HBV1.3 DNA via hydrodynamic injection or infected with 1×10 11 vg of AAV/HBV ( n =3). ( b ) ALT levels of HBV carrier mice were measured over time ( n =3). ( c ) H E staining of liver sections from mice infected with different doses of AAV/HBV. One of two independent experiments is presented. AAV, adeno-associated virus; ALT, alanine aminotransferase; HBsAg, HBV surface antigen; HBV, hepatitis B virus; H E, hematoxylin and eosin.

    Journal: Cellular and Molecular Immunology

    Article Title: A mouse model for HBV immunotolerance and immunotherapy

    doi: 10.1038/cmi.2013.43

    Figure Lengend Snippet: The immune response was undetectable in AAV/HBV-infected mice. ( a ) Serum HBsAb levels in C57BL/6 mice receiving 15 µg of pBlue-HBV1.3 DNA via hydrodynamic injection or infected with 1×10 11 vg of AAV/HBV ( n =3). ( b ) ALT levels of HBV carrier mice were measured over time ( n =3). ( c ) H E staining of liver sections from mice infected with different doses of AAV/HBV. One of two independent experiments is presented. AAV, adeno-associated virus; ALT, alanine aminotransferase; HBsAg, HBV surface antigen; HBV, hepatitis B virus; H E, hematoxylin and eosin.

    Article Snippet: Liver HBV DNA was extracted from 50 mg of liver tissues using a genomic DNA kit (TIANGEN Biotech, Beijing, China).

    Techniques: Infection, Mouse Assay, Injection, Staining

    Amplification and analysis of the nifH gene of SnebYK. (A) Amplicon of SnebYK nifH (363 bp) in the left lane and DNA ladder in the right lane. (B) Transcriptional analysis of the SnebYK nifH gene in nitrogen-free medium determined with RT-PCR. The lane labeled “-N” shows the expression of the 16S rRNA and nifH gene of SnebYK grown in ACCC55 nitrogen-free medium; the lane labeled “+N” shows the expression of the 16S rRNA and nifH gene of SnebYK grown in nutrient agar medium. The transcript level of the 16S rRNA was used as a loading control; the transcript level of nifH of SnebYK grown in nutrient agar medium was used as a negative control. (C) Dendrogram based on the nifH sequences of SnebYK and other strains in the genus Klebsiella with similar nifH sequences. This analysis was performed using the neighbor-joining method in Mega 7.0.26 with a bootstrap value of n = 1000.

    Journal: Frontiers in Microbiology

    Article Title: Klebsiella pneumoniae SnebYK Mediates Resistance Against Heterodera glycines and Promotes Soybean Growth

    doi: 10.3389/fmicb.2018.01134

    Figure Lengend Snippet: Amplification and analysis of the nifH gene of SnebYK. (A) Amplicon of SnebYK nifH (363 bp) in the left lane and DNA ladder in the right lane. (B) Transcriptional analysis of the SnebYK nifH gene in nitrogen-free medium determined with RT-PCR. The lane labeled “-N” shows the expression of the 16S rRNA and nifH gene of SnebYK grown in ACCC55 nitrogen-free medium; the lane labeled “+N” shows the expression of the 16S rRNA and nifH gene of SnebYK grown in nutrient agar medium. The transcript level of the 16S rRNA was used as a loading control; the transcript level of nifH of SnebYK grown in nutrient agar medium was used as a negative control. (C) Dendrogram based on the nifH sequences of SnebYK and other strains in the genus Klebsiella with similar nifH sequences. This analysis was performed using the neighbor-joining method in Mega 7.0.26 with a bootstrap value of n = 1000.

    Article Snippet: To analyze the nifH gene of SnebYK, genomic DNA was extracted from the SnebYK strain using a bacterial genomic DNA extraction kit (Tiangen Biotech, China).

    Techniques: Amplification, Reverse Transcription Polymerase Chain Reaction, Labeling, Expressing, Negative Control

    Effects of Lfcin4 and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine (DNA) (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).

    Journal: Scientific Reports

    Article Title: Killing of Staphylococcus aureus and Salmonella enteritidis and neutralization of lipopolysaccharide by 17-residue bovine lactoferricins: improved activity of Trp/Ala-containing molecules

    doi: 10.1038/srep44278

    Figure Lengend Snippet: Effects of Lfcin4 and Lfcin5 on macromolecular synthesis in S. aureus ATCC25923. Incorporation of 3 H-glucosamine hydrochloride (peptidoglycan) (A) , 3 H-leucine (protein) (B) , 3 H-thymidine (DNA) (C) , and 3 H-uridine (RNA) (D) was determined in cells treated with 1 × MIC Lfcin4 and Lfcin5. Van: vancomycin (2 × MIC); Ery: erythromycin (2 × MIC); Cip: ciprofloxacin (8 × MIC); Rif: rifampicin (4 × MIC). Antibiotics were used as controls. Results are presented as the mean ± SEM (n = 3).

    Article Snippet: Effects of Lfcin4 and Lfcin5 on bacterial genomic DNA Genomic DNA was extracted from S. aureus ATCC25923, E. coli O157 and S. enteritidis CVCC3377 using a TIANamp Bacteria DNA Kit (TIANGEN Biotech Co., Ltd., Beijing).

    Techniques:

    In vitro binding of Lfcin4 and Lfcin5 to bacterial genomic DNA. (A) Gel retardation analysis of the binding of Lfcin4 and Lfcin5 to genomic DNA. M: DNA marker λDNA/ Hind III. Lanes 1–6: genomic DNA from E. coli CICC21530; Lanes 7–12: genomic DNA from S. aureus ATCC25923; Lanes 13–18: genomic DNA from S. enteritidis CVCC3377. The mass ratios of peptide to genomic DNA were 10, 5, 2.5, 1, 0.5, and 0. Full-length gels are presented in Supplementary Figure S2 . (B–D) CD spectra of genomic DNA from E. coli CICC21530 (B) , S. aureus ATCC25923 (C) and S. enteritidis CVCC3377 (D) in the presence of Lfcin4 and Lfcin5. The mass ratios of peptide to DNA were 5 and 10.

    Journal: Scientific Reports

    Article Title: Killing of Staphylococcus aureus and Salmonella enteritidis and neutralization of lipopolysaccharide by 17-residue bovine lactoferricins: improved activity of Trp/Ala-containing molecules

    doi: 10.1038/srep44278

    Figure Lengend Snippet: In vitro binding of Lfcin4 and Lfcin5 to bacterial genomic DNA. (A) Gel retardation analysis of the binding of Lfcin4 and Lfcin5 to genomic DNA. M: DNA marker λDNA/ Hind III. Lanes 1–6: genomic DNA from E. coli CICC21530; Lanes 7–12: genomic DNA from S. aureus ATCC25923; Lanes 13–18: genomic DNA from S. enteritidis CVCC3377. The mass ratios of peptide to genomic DNA were 10, 5, 2.5, 1, 0.5, and 0. Full-length gels are presented in Supplementary Figure S2 . (B–D) CD spectra of genomic DNA from E. coli CICC21530 (B) , S. aureus ATCC25923 (C) and S. enteritidis CVCC3377 (D) in the presence of Lfcin4 and Lfcin5. The mass ratios of peptide to DNA were 5 and 10.

    Article Snippet: Effects of Lfcin4 and Lfcin5 on bacterial genomic DNA Genomic DNA was extracted from S. aureus ATCC25923, E. coli O157 and S. enteritidis CVCC3377 using a TIANamp Bacteria DNA Kit (TIANGEN Biotech Co., Ltd., Beijing).

    Techniques: In Vitro, Binding Assay, Electrophoretic Mobility Shift Assay, Marker

    Growth of Hsp90 knockout parasite ( ΔHsp90 ) in vitro. The parasites were cultured in African green monkey kidney (Vero) cells, 10 5 T. gondii were added to the 6-well plates, and the infection ratio was 1:1. Observation of RH Δku80 (A), ΔHsp90 (B), and complemented (C) parasites by inverted microscope. The plaque produced by ΔPKAR strains (Fig. 6B) was significantly smaller than that of RH Δku80 and complemented parasites (Figs. 6A, 6C), scale bar = 20 μm. T. gondii tachyzoites and Vero cells were indicated by arrows, T. gondii (black arrow), Vero cells (white arrow). (D) The parasites were collected at the same time, and genomic DNA was extracted by TIANGEN kit. T. gondii DNA was detected by SYBR-green real-time PCR using B1 primer pairs, the standard curve was obtained by the known concentration of the RH Δku80 parasites with the primers (B1), and the parasite number was calculated by interpolation from this standard curve. ** p

    Journal: Parasite

    Article Title: The heat shock protein 90 of Toxoplasma gondii is essential for invasion of host cells and tachyzoite growth

    doi: 10.1051/parasite/2017023

    Figure Lengend Snippet: Growth of Hsp90 knockout parasite ( ΔHsp90 ) in vitro. The parasites were cultured in African green monkey kidney (Vero) cells, 10 5 T. gondii were added to the 6-well plates, and the infection ratio was 1:1. Observation of RH Δku80 (A), ΔHsp90 (B), and complemented (C) parasites by inverted microscope. The plaque produced by ΔPKAR strains (Fig. 6B) was significantly smaller than that of RH Δku80 and complemented parasites (Figs. 6A, 6C), scale bar = 20 μm. T. gondii tachyzoites and Vero cells were indicated by arrows, T. gondii (black arrow), Vero cells (white arrow). (D) The parasites were collected at the same time, and genomic DNA was extracted by TIANGEN kit. T. gondii DNA was detected by SYBR-green real-time PCR using B1 primer pairs, the standard curve was obtained by the known concentration of the RH Δku80 parasites with the primers (B1), and the parasite number was calculated by interpolation from this standard curve. ** p

    Article Snippet: Next, 24, 48, 72, and 96 h post-infection (PI), the parasites were collected and genomic DNA was extracted using the TIANGEN genomic DNA isolation kit by following the manufacturer’s protocol (TIANGEN, Beijing).

    Techniques: Knock-Out, In Vitro, Cell Culture, Infection, Inverted Microscopy, Produced, SYBR Green Assay, Real-time Polymerase Chain Reaction, Concentration Assay

    TALENs-mediated homozygous recombination. (A) T7E1 enzyme analysis of transfected CD4 + U87 cells after each transfection with CCR5-TALENs. The targeting efficiencies after first, second and third rounds of transfection were 14.80, 38.20 and 50.04%, respectively. M, 100 bp marker; wild-type, wild-type cells; lanes 1–3, transfected cells after the first, second and third rounds of transfections. (B) PCR of CCR5Δ32/Δ1 genotype cells. M, 100 bp marker; lane 1, prior to transfection; lane 2, after CCR5-TALENs transfection; lane 3, after CCR5Δ1-TALENs transfection; the CCR5Δ32 band (168 bp) was 2.87-times brighter than the CCR5Δ1 band; lane 4, wild-type cells (negative control); lane 5, CCR5Δ32 donor DNA (positive control). (C) Sequencing of wild-type CD4 + U87 cells and those with CCR5Δ32/Δ1 mutations. (D) PCR of 34 single-cell cultured clones of CD4 + U87 cells with CCR5Δ32/Δ1 mutations post-transfection with CCR5Δ1-TALENs. The CCR5Δ32 band was observed in all clones; however, as a single band, it was only seen in clones 16, 18 and 25 post-transfection with CCR5Δ1-TALENs. M, 100 bp marker; lanes 1–34, single-cell cultured cells post-transfection with CCR5Δ1-TALENs; wild-type, wild-type cells (negative control); donor, CCR5Δ32 donor DNA (positive control). (E) Gene sequencing of clones 16, 18 and 25. CCR5, C-C chemokine receptor type 5; CD4, cluster of differentiation 4; PCR, polymerase chain reaction; T7E1, T7 endonuclease 1; TALENs, transcription activator-like effector nucleases.

    Journal: Molecular Medicine Reports

    Article Title: TALENs-mediated homozygous CCR5Δ32 mutations endow CD4+ U87 cells with resistance against HIV-1 infection

    doi: 10.3892/mmr.2017.7889

    Figure Lengend Snippet: TALENs-mediated homozygous recombination. (A) T7E1 enzyme analysis of transfected CD4 + U87 cells after each transfection with CCR5-TALENs. The targeting efficiencies after first, second and third rounds of transfection were 14.80, 38.20 and 50.04%, respectively. M, 100 bp marker; wild-type, wild-type cells; lanes 1–3, transfected cells after the first, second and third rounds of transfections. (B) PCR of CCR5Δ32/Δ1 genotype cells. M, 100 bp marker; lane 1, prior to transfection; lane 2, after CCR5-TALENs transfection; lane 3, after CCR5Δ1-TALENs transfection; the CCR5Δ32 band (168 bp) was 2.87-times brighter than the CCR5Δ1 band; lane 4, wild-type cells (negative control); lane 5, CCR5Δ32 donor DNA (positive control). (C) Sequencing of wild-type CD4 + U87 cells and those with CCR5Δ32/Δ1 mutations. (D) PCR of 34 single-cell cultured clones of CD4 + U87 cells with CCR5Δ32/Δ1 mutations post-transfection with CCR5Δ1-TALENs. The CCR5Δ32 band was observed in all clones; however, as a single band, it was only seen in clones 16, 18 and 25 post-transfection with CCR5Δ1-TALENs. M, 100 bp marker; lanes 1–34, single-cell cultured cells post-transfection with CCR5Δ1-TALENs; wild-type, wild-type cells (negative control); donor, CCR5Δ32 donor DNA (positive control). (E) Gene sequencing of clones 16, 18 and 25. CCR5, C-C chemokine receptor type 5; CD4, cluster of differentiation 4; PCR, polymerase chain reaction; T7E1, T7 endonuclease 1; TALENs, transcription activator-like effector nucleases.

    Article Snippet: After three rounds of transfection, DNA was extracted using the Blood Genomic DNA Extraction Mini kit (Tiangen Biotech Co., Ltd., Beijing, China) for T7 endonuclease 1 (T7E1) enzyme analysis, and ≥50 transfected cells were cultured individually.

    Techniques: TALENs, Transfection, Marker, Polymerase Chain Reaction, Negative Control, Positive Control, Sequencing, Cell Culture, Clone Assay