Structured Review

iNtRON Biotechnology cdna template
Gene expression of cytosine deaminase (CD), rabbit carboxyl esterase (CE) and interferon-beta (IFN-β) in the genetically engineered stem cells. ( A ) <t>cDNA</t> was produced by semi-quantitative <t>RT-PCR</t> to confirm the expression of CE, CD, and/or IFN-β genes; ( B ) Relative expression levels of CD gene in HB1.F3, HB1.F3.CD, and HB1.F3.CD.IFN-β cells were confirmed with quantitative real-time PCR (qRT-PCR). Mwt, molecular weight; Negative control, without cDNA template.
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1) Product Images from "Effects of Genetically Engineered Stem Cells Expressing Cytosine Deaminase and Interferon-Beta or Carboxyl Esterase on the Growth of LNCaP Prostate Cancer Cells"

Article Title: Effects of Genetically Engineered Stem Cells Expressing Cytosine Deaminase and Interferon-Beta or Carboxyl Esterase on the Growth of LNCaP Prostate Cancer Cells

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms131012519

Gene expression of cytosine deaminase (CD), rabbit carboxyl esterase (CE) and interferon-beta (IFN-β) in the genetically engineered stem cells. ( A ) cDNA was produced by semi-quantitative RT-PCR to confirm the expression of CE, CD, and/or IFN-β genes; ( B ) Relative expression levels of CD gene in HB1.F3, HB1.F3.CD, and HB1.F3.CD.IFN-β cells were confirmed with quantitative real-time PCR (qRT-PCR). Mwt, molecular weight; Negative control, without cDNA template.
Figure Legend Snippet: Gene expression of cytosine deaminase (CD), rabbit carboxyl esterase (CE) and interferon-beta (IFN-β) in the genetically engineered stem cells. ( A ) cDNA was produced by semi-quantitative RT-PCR to confirm the expression of CE, CD, and/or IFN-β genes; ( B ) Relative expression levels of CD gene in HB1.F3, HB1.F3.CD, and HB1.F3.CD.IFN-β cells were confirmed with quantitative real-time PCR (qRT-PCR). Mwt, molecular weight; Negative control, without cDNA template.

Techniques Used: Expressing, Produced, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Molecular Weight, Negative Control

2) Product Images from "Cell Growth of BG-1 Ovarian Cancer Cells was Promoted by 4-Tert-octylphenol and 4-Nonylphenol via Downregulation of TGF-? Receptor 2 and Upregulation of c-myc"

Article Title: Cell Growth of BG-1 Ovarian Cancer Cells was Promoted by 4-Tert-octylphenol and 4-Nonylphenol via Downregulation of TGF-? Receptor 2 and Upregulation of c-myc

Journal: Toxicological Research

doi: 10.5487/TR.2011.27.4.253

Altered expression levels of TGF-βR1 gene and TGF-βR2 gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated with E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). ( A ) Expression levels of TGF-βR1 by treatment with E2, OP or NP. ( B ) Expression levels of TGF-βR2 by treatment with E2, OP or NP. Expression level of TGF-βR1 and TGF-βR2 was detected by using semi-quantitative reverse-transcription PCR. Data represent the means ± S.D. of triplicate experiments. *, p
Figure Legend Snippet: Altered expression levels of TGF-βR1 gene and TGF-βR2 gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated with E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). ( A ) Expression levels of TGF-βR1 by treatment with E2, OP or NP. ( B ) Expression levels of TGF-βR2 by treatment with E2, OP or NP. Expression level of TGF-βR1 and TGF-βR2 was detected by using semi-quantitative reverse-transcription PCR. Data represent the means ± S.D. of triplicate experiments. *, p

Techniques Used: Expressing, Polymerase Chain Reaction

Altered expression levels of TGF-β1 gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated with E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). Expression level of TGF-β1 was detected by using semi-quantitative reverse-transcription PCR. PCR products were run on a 1.5% agarose gel, bands were scanned and the density of the bands on the gel was quantified using Gel Doc 2000 as described in Materials and Methods. Data represent the means ± S.D. of triplicate experiments. *, p
Figure Legend Snippet: Altered expression levels of TGF-β1 gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated with E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). Expression level of TGF-β1 was detected by using semi-quantitative reverse-transcription PCR. PCR products were run on a 1.5% agarose gel, bands were scanned and the density of the bands on the gel was quantified using Gel Doc 2000 as described in Materials and Methods. Data represent the means ± S.D. of triplicate experiments. *, p

Techniques Used: Expressing, Polymerase Chain Reaction, Agarose Gel Electrophoresis

Altered expression levels of c-myc gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). Expression level of c-myc was detected by using semi-quantitative reverse-transcription PCR. Data represent the means ± S.D. of triplicate experiments. *, p
Figure Legend Snippet: Altered expression levels of c-myc gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). Expression level of c-myc was detected by using semi-quantitative reverse-transcription PCR. Data represent the means ± S.D. of triplicate experiments. *, p

Techniques Used: Expressing, Polymerase Chain Reaction

3) Product Images from "Molecular and immunological characterization of Hyalomma dromedarii and Hyalomma excavatum (Acari: Ixodidae) vectors of Q fever in camels"

Article Title: Molecular and immunological characterization of Hyalomma dromedarii and Hyalomma excavatum (Acari: Ixodidae) vectors of Q fever in camels

Journal: Veterinary World

doi: 10.14202/vetworld.2018.1109-1119

Agarose gel electrophoresis of polymerase chain reaction products obtained from Hyalomma dromedarii and Hyalomma excavatum using two genes: (a) 16S ribosomal RNA gene reveals 440 bp, (b) cytochrome oxidase subunit-1 gene, reveals 850 bp, (M) 1 kb DNA ladder, (N) negative control.
Figure Legend Snippet: Agarose gel electrophoresis of polymerase chain reaction products obtained from Hyalomma dromedarii and Hyalomma excavatum using two genes: (a) 16S ribosomal RNA gene reveals 440 bp, (b) cytochrome oxidase subunit-1 gene, reveals 850 bp, (M) 1 kb DNA ladder, (N) negative control.

Techniques Used: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Negative Control

Agarose gel electrophoresis of polymerase chain reaction products obtained from H. dromedarii using IS30A spacer: (M) 1 kb DNA ladder, (N) negative control, lanes 7 and 8 represent positive tick samples and lanes 1-6 represent negative tick samples.
Figure Legend Snippet: Agarose gel electrophoresis of polymerase chain reaction products obtained from H. dromedarii using IS30A spacer: (M) 1 kb DNA ladder, (N) negative control, lanes 7 and 8 represent positive tick samples and lanes 1-6 represent negative tick samples.

Techniques Used: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Negative Control

4) Product Images from "Inhibition of Lipopolysaccharide-Induced Proinflammatory Responses by Buddleja officinalis Extract in BV-2 Microglial Cells via Negative Regulation of NF-κB and ERK1/2 Signaling"

Article Title: Inhibition of Lipopolysaccharide-Induced Proinflammatory Responses by Buddleja officinalis Extract in BV-2 Microglial Cells via Negative Regulation of NF-κB and ERK1/2 Signaling

Journal: Molecules

doi: 10.3390/molecules18089195

Effect of BOWE on LPS-induced mRNA expression levels of iNOS and COX-2 in BV-2 cells: For mRNA expression, BV-2 cells were pretreated with BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated, and mRNA levels of iNOS (A) and COX-2 (B) were then measured by RT-PCR. β actin expression was used as an internal control. The quantification of relative band intensities from three independent experiments were analyzed by densitometry and expressed as means ± SEM. * p
Figure Legend Snippet: Effect of BOWE on LPS-induced mRNA expression levels of iNOS and COX-2 in BV-2 cells: For mRNA expression, BV-2 cells were pretreated with BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated, and mRNA levels of iNOS (A) and COX-2 (B) were then measured by RT-PCR. β actin expression was used as an internal control. The quantification of relative band intensities from three independent experiments were analyzed by densitometry and expressed as means ± SEM. * p

Techniques Used: Expressing, Isolation, Reverse Transcription Polymerase Chain Reaction

Effect of BOWE on the mRNA expressions of IL-1β ( A ), IL-6 ( B ), and TNF-α ( C ): BV-2 cells were pretreated with or without the indicated concentrations of BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated and mRNA expressions of the aforementioned cytokines were measured by RT-PCR. β actin was used as a housekeeping gene. The quantification of relative band intensities from three independentexperiments were analyzed by densitometry and expressed as means ± SEM. * p
Figure Legend Snippet: Effect of BOWE on the mRNA expressions of IL-1β ( A ), IL-6 ( B ), and TNF-α ( C ): BV-2 cells were pretreated with or without the indicated concentrations of BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated and mRNA expressions of the aforementioned cytokines were measured by RT-PCR. β actin was used as a housekeeping gene. The quantification of relative band intensities from three independentexperiments were analyzed by densitometry and expressed as means ± SEM. * p

Techniques Used: Isolation, Reverse Transcription Polymerase Chain Reaction

5) Product Images from "Inhibition of Lipopolysaccharide-Induced Proinflammatory Responses by Buddleja officinalis Extract in BV-2 Microglial Cells via Negative Regulation of NF-κB and ERK1/2 Signaling"

Article Title: Inhibition of Lipopolysaccharide-Induced Proinflammatory Responses by Buddleja officinalis Extract in BV-2 Microglial Cells via Negative Regulation of NF-κB and ERK1/2 Signaling

Journal: Molecules

doi: 10.3390/molecules18089195

Effect of BOWE on LPS-induced mRNA expression levels of iNOS and COX-2 in BV-2 cells: For mRNA expression, BV-2 cells were pretreated with BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated, and mRNA levels of iNOS (A) and COX-2 (B) were then measured by RT-PCR. β actin expression was used as an internal control. The quantification of relative band intensities from three independent experiments were analyzed by densitometry and expressed as means ± SEM. * p
Figure Legend Snippet: Effect of BOWE on LPS-induced mRNA expression levels of iNOS and COX-2 in BV-2 cells: For mRNA expression, BV-2 cells were pretreated with BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated, and mRNA levels of iNOS (A) and COX-2 (B) were then measured by RT-PCR. β actin expression was used as an internal control. The quantification of relative band intensities from three independent experiments were analyzed by densitometry and expressed as means ± SEM. * p

Techniques Used: Expressing, Isolation, Reverse Transcription Polymerase Chain Reaction

Effect of BOWE on the mRNA expressions of IL-1β ( A ), IL-6 ( B ), and TNF-α ( C ): BV-2 cells were pretreated with or without the indicated concentrations of BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated and mRNA expressions of the aforementioned cytokines were measured by RT-PCR. β actin was used as a housekeeping gene. The quantification of relative band intensities from three independentexperiments were analyzed by densitometry and expressed as means ± SEM. * p
Figure Legend Snippet: Effect of BOWE on the mRNA expressions of IL-1β ( A ), IL-6 ( B ), and TNF-α ( C ): BV-2 cells were pretreated with or without the indicated concentrations of BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated and mRNA expressions of the aforementioned cytokines were measured by RT-PCR. β actin was used as a housekeeping gene. The quantification of relative band intensities from three independentexperiments were analyzed by densitometry and expressed as means ± SEM. * p

Techniques Used: Isolation, Reverse Transcription Polymerase Chain Reaction

6) Product Images from "Molecular cloning, purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein"

Article Title: Molecular cloning, purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein

Journal: Journal of Veterinary Science

doi: 10.4142/jvs.2016.17.1.119

Amplification of Brucella ( B. ) abortus mdh gene. PCR products on 1.5% (w/v) agarose gel. Lane 1, DNA marker; Lane 2, single expected band of mdh gene (963 bp, arrow).
Figure Legend Snippet: Amplification of Brucella ( B. ) abortus mdh gene. PCR products on 1.5% (w/v) agarose gel. Lane 1, DNA marker; Lane 2, single expected band of mdh gene (963 bp, arrow).

Techniques Used: Amplification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Marker

7) Product Images from "Core binding factor β of osteoblasts maintains cortical bone mass via stabilization of Runx2 in mice"

Article Title: Core binding factor β of osteoblasts maintains cortical bone mass via stabilization of Runx2 in mice

Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

doi: 10.1002/jbmr.2397

Osteoblast-specific deletion of Cbfb by Col1a1-Cre The osteoblast-specific deletion of Cbfb was determined by PCR using genomic DNA obtained from tibia (A). Reduction of Cbfβ was confirmed by Q-PCR using RNA isolated from bone tissue of 4-week-old mice (B). Immunohistochemistry of tibia shows deletion of Cbfβ in Cbfb ∆ob/∆ob mice (C). The tibiae of 12-week-old Cbfb Δob/Δob mice were used (A and C). WT mice (□, n = 3), Cbfb Δob/Δob mice (■, n = 3). ** p
Figure Legend Snippet: Osteoblast-specific deletion of Cbfb by Col1a1-Cre The osteoblast-specific deletion of Cbfb was determined by PCR using genomic DNA obtained from tibia (A). Reduction of Cbfβ was confirmed by Q-PCR using RNA isolated from bone tissue of 4-week-old mice (B). Immunohistochemistry of tibia shows deletion of Cbfβ in Cbfb ∆ob/∆ob mice (C). The tibiae of 12-week-old Cbfb Δob/Δob mice were used (A and C). WT mice (□, n = 3), Cbfb Δob/Δob mice (■, n = 3). ** p

Techniques Used: Polymerase Chain Reaction, Isolation, Mouse Assay, Immunohistochemistry

8) Product Images from "Molecular cloning, purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein"

Article Title: Molecular cloning, purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein

Journal: Journal of Veterinary Science

doi: 10.4142/jvs.2016.17.1.119

Amplification of Brucella ( B. ) abortus mdh gene. PCR products on 1.5% (w/v) agarose gel. Lane 1, DNA marker; Lane 2, single expected band of mdh gene (963 bp, arrow).
Figure Legend Snippet: Amplification of Brucella ( B. ) abortus mdh gene. PCR products on 1.5% (w/v) agarose gel. Lane 1, DNA marker; Lane 2, single expected band of mdh gene (963 bp, arrow).

Techniques Used: Amplification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Marker

9) Product Images from "In Vitro Characterization of Lactobacillus plantarum Strains with Inhibitory Activity on Enteropathogens for Use as Potential Animal Probiotics"

Article Title: In Vitro Characterization of Lactobacillus plantarum Strains with Inhibitory Activity on Enteropathogens for Use as Potential Animal Probiotics

Journal: Indian Journal of Microbiology

doi: 10.1007/s12088-017-0646-4

Identification of strains MJM60319, MJM60298, and MJM60399. a Morphology of strains MJM60319, MJM60298, and MJM60399 observed by scanning electron microscopy. Scale bar indicates 1 µm, b phylogenetic tree constructed based on 16S rDNA sequence of the strain. The strains MJM60319, MJM60298, and MJM60399 possesses 99% similarity with L. plantarum . c GTG5 PCR fingerprinting analysis of LAB strains. 1, MJM60319; 2, MJM60298; 3, MJM60399; 4, L. plantarum ssp. plantarum KACC 11451; 5, L. paraplantarum KACC 12373; 6, L. pentosus KACC 12428; M, Marker: 100–10,000 bp GeneRulerTM DNA ladder mix (Fermentas)
Figure Legend Snippet: Identification of strains MJM60319, MJM60298, and MJM60399. a Morphology of strains MJM60319, MJM60298, and MJM60399 observed by scanning electron microscopy. Scale bar indicates 1 µm, b phylogenetic tree constructed based on 16S rDNA sequence of the strain. The strains MJM60319, MJM60298, and MJM60399 possesses 99% similarity with L. plantarum . c GTG5 PCR fingerprinting analysis of LAB strains. 1, MJM60319; 2, MJM60298; 3, MJM60399; 4, L. plantarum ssp. plantarum KACC 11451; 5, L. paraplantarum KACC 12373; 6, L. pentosus KACC 12428; M, Marker: 100–10,000 bp GeneRulerTM DNA ladder mix (Fermentas)

Techniques Used: Electron Microscopy, Construct, Sequencing, Polymerase Chain Reaction, Marker

10) Product Images from "Torilin Inhibits Inflammation by Limiting TAK1-Mediated MAP Kinase and NF-κB Activation"

Article Title: Torilin Inhibits Inflammation by Limiting TAK1-Mediated MAP Kinase and NF-κB Activation

Journal: Mediators of Inflammation

doi: 10.1155/2017/7250968

Torilin restores I- κ B α phosphorylation and degradation; thereby it inhibits p65-NF- κ B nuclear translocation. (a) RAW 264.7 cells were pretreated with the indicated concentrations of torilin for 30 min and incubated with LPS (100 ng/ml) for (15–60 min) time course and then assayed for the phosphorylation and degradation of I- κ B α and nuclear translocation of p65 by western immunoblot analysis as described under Materials and Methods. (b and c) RAW macrophages were treated with LPS (100 ngmL −1 ) and 25 µ M torilin, respectively, for the indicated time course before cytoplasmic and nuclear protein fractions were subjected to western blot analyses for p65 (upper panel) and p50 (middle panel) of cytoplasmic (b) and nuclear (c) proteins, respectively. β -Actin and ploy(ADP-ribose) polymerase (PARP) were used as a control for the cytoplasmic and nuclear protein loading, respectively. Images are representative of 3 or 4 independent experiments. Values in bar graphs are means ± SE of at least 4 independent experiments performed in triplicate. Significance was determined using Student's t- test. ∗ P
Figure Legend Snippet: Torilin restores I- κ B α phosphorylation and degradation; thereby it inhibits p65-NF- κ B nuclear translocation. (a) RAW 264.7 cells were pretreated with the indicated concentrations of torilin for 30 min and incubated with LPS (100 ng/ml) for (15–60 min) time course and then assayed for the phosphorylation and degradation of I- κ B α and nuclear translocation of p65 by western immunoblot analysis as described under Materials and Methods. (b and c) RAW macrophages were treated with LPS (100 ngmL −1 ) and 25 µ M torilin, respectively, for the indicated time course before cytoplasmic and nuclear protein fractions were subjected to western blot analyses for p65 (upper panel) and p50 (middle panel) of cytoplasmic (b) and nuclear (c) proteins, respectively. β -Actin and ploy(ADP-ribose) polymerase (PARP) were used as a control for the cytoplasmic and nuclear protein loading, respectively. Images are representative of 3 or 4 independent experiments. Values in bar graphs are means ± SE of at least 4 independent experiments performed in triplicate. Significance was determined using Student's t- test. ∗ P

Techniques Used: Translocation Assay, Incubation, Western Blot

Torilin arrests AP-1 and NF- κ B DNA binding and reporter gene expression in LPS-stimulated RAW 264.7 cells. RAW macrophages were treated with vehicle or torilin (as indicated) and stimulated with LPS for 45 min before nuclear protein was isolated. DNA binding was analyzed using specific γ 32P-labeled oligonucleotide probes for NF- κ B (a) or AP-1 (b). Specificity was demonstrated by coincubation with a 25-fold excess of unlabeled specific probe of NF- κ B and AP-1 for competition. Cells were transiently transfected with NF- κ B (c) or AP-1 (d), plasmids treated with the indicated concentrations of torilin and LPS (100 ngmL −1 ) for 6 h and assayed for CAT expression using a CAT enzyme-linked immunosorbent assay kit. Each column shows the mean ± SEM of quadruplicate determinations. Luciferase activity was normalized to β -galactosidase activity. Images are representative of 3 independent experiments. ∗ P
Figure Legend Snippet: Torilin arrests AP-1 and NF- κ B DNA binding and reporter gene expression in LPS-stimulated RAW 264.7 cells. RAW macrophages were treated with vehicle or torilin (as indicated) and stimulated with LPS for 45 min before nuclear protein was isolated. DNA binding was analyzed using specific γ 32P-labeled oligonucleotide probes for NF- κ B (a) or AP-1 (b). Specificity was demonstrated by coincubation with a 25-fold excess of unlabeled specific probe of NF- κ B and AP-1 for competition. Cells were transiently transfected with NF- κ B (c) or AP-1 (d), plasmids treated with the indicated concentrations of torilin and LPS (100 ngmL −1 ) for 6 h and assayed for CAT expression using a CAT enzyme-linked immunosorbent assay kit. Each column shows the mean ± SEM of quadruplicate determinations. Luciferase activity was normalized to β -galactosidase activity. Images are representative of 3 independent experiments. ∗ P

Techniques Used: Binding Assay, Expressing, Isolation, Labeling, Transfection, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay

Torilin pretreatment reduces LPS-induced proinflammatory cytokine secretions into the culture media. RAW 264.7 cells were pretreated with torilin or vehicle for 30 min and stimulated with LPS for 24 h. TNF- α (a), IL-1 β (b), IL-6 (c), and GM-CSF (d) were determined using ELISA. Each bar graph represents mean ± (SE) for four independent experiments. Significance was determined using Student's t -test. ∗ P
Figure Legend Snippet: Torilin pretreatment reduces LPS-induced proinflammatory cytokine secretions into the culture media. RAW 264.7 cells were pretreated with torilin or vehicle for 30 min and stimulated with LPS for 24 h. TNF- α (a), IL-1 β (b), IL-6 (c), and GM-CSF (d) were determined using ELISA. Each bar graph represents mean ± (SE) for four independent experiments. Significance was determined using Student's t -test. ∗ P

Techniques Used: Enzyme-linked Immunosorbent Assay

Effect of torilin on mitogen activated protein kinase (MAPK) activation. LPS-stimulated RAW 264.7 macrophages were treated with vehicle or torilin (6.25–25 µ M) and stimulated with LPS for the indicated periods of time. Total protein was subjected to western blot analyses using total and phospho-anti-RK1/2, p38 MAPK , and JNK1/2 antibodies. Torilin suppressed phosphorylation of the three MAPKs (a and b). Cells were further pretreated with vehicle or PD98059 (30 µ M), ERK1/2 inhibitor, SB203580 (10 µ M), p38 MAPK inhibitor, or SP600125 (10 µ M), JNK1/2 inhibitor, with torilin and stimulated with 100 ngmL −1 LPS as indicated in (c). Significance was determined using Student's t- test versus the control group. Images are representative of 3 or 4 independent experiments. Values in bar graphs are means ± SE of at least 4 independent experiments performed in triplicate. ∗ P
Figure Legend Snippet: Effect of torilin on mitogen activated protein kinase (MAPK) activation. LPS-stimulated RAW 264.7 macrophages were treated with vehicle or torilin (6.25–25 µ M) and stimulated with LPS for the indicated periods of time. Total protein was subjected to western blot analyses using total and phospho-anti-RK1/2, p38 MAPK , and JNK1/2 antibodies. Torilin suppressed phosphorylation of the three MAPKs (a and b). Cells were further pretreated with vehicle or PD98059 (30 µ M), ERK1/2 inhibitor, SB203580 (10 µ M), p38 MAPK inhibitor, or SP600125 (10 µ M), JNK1/2 inhibitor, with torilin and stimulated with 100 ngmL −1 LPS as indicated in (c). Significance was determined using Student's t- test versus the control group. Images are representative of 3 or 4 independent experiments. Values in bar graphs are means ± SE of at least 4 independent experiments performed in triplicate. ∗ P

Techniques Used: Activation Assay, Western Blot

Torilin inhibits LPS-induced NO release, PGE2 secretion, and protein as well as mRNA expression of iNOS and COX-2 enzymes. RAW 246.7 macrophages were pretreated with torilin or vehicle for 30 min and stimulated with LPS for 18 or 24 h. (a) Cell culture supernatants were analyzed for nitrite release, as a measure of NO production. (b) PGE 2 secretion in culture media was analyzed in torilin treated RAW cells as described in Materials and Methods. After 24 h of stimulation for protein expression (c) and 18 h of stimulation for mRNA expression (d) were depicted, respectively. Again, COX-2 protein (e) and mRNA (f) were determined by western blot and RT-PCR, respectively. GAPDH and β -actin were used as controls for mRNA and protein loading, respectively. Images are representative of 3 or 4 independent experiments. Values in bar graphs are means ± SE of at least 4 independent experiments performed in triplicate. Significance was determined using Student's t- test versus the control group. ∗ P
Figure Legend Snippet: Torilin inhibits LPS-induced NO release, PGE2 secretion, and protein as well as mRNA expression of iNOS and COX-2 enzymes. RAW 246.7 macrophages were pretreated with torilin or vehicle for 30 min and stimulated with LPS for 18 or 24 h. (a) Cell culture supernatants were analyzed for nitrite release, as a measure of NO production. (b) PGE 2 secretion in culture media was analyzed in torilin treated RAW cells as described in Materials and Methods. After 24 h of stimulation for protein expression (c) and 18 h of stimulation for mRNA expression (d) were depicted, respectively. Again, COX-2 protein (e) and mRNA (f) were determined by western blot and RT-PCR, respectively. GAPDH and β -actin were used as controls for mRNA and protein loading, respectively. Images are representative of 3 or 4 independent experiments. Values in bar graphs are means ± SE of at least 4 independent experiments performed in triplicate. Significance was determined using Student's t- test versus the control group. ∗ P

Techniques Used: Expressing, Cell Culture, Western Blot, Reverse Transcription Polymerase Chain Reaction

Torilin suppresses LPS-induced IKK and MAP3K complex activation and inhibits the TAK1 interaction with the components of MAPK and NF- κ B signaling pathways. RAW macrophage cells were incubated with torilin or vehicle for 30 min and then stimulated with LPS as indicated in the figures. (a) Lysates from torilin or vehicle treated-cells were immunoblotted to detect the activation status of IKK. (b) Immunoblots depicting the time dependent effects of torilin in TAK1, MKK4, and IKK phosphorylation. (c) The kinase activity of the TAK1 was determined after immunoprecipitates were resuspended in kinase buffer and assayed for kinase activity as described in Materials and Methods. (d) Cell lysates were immunoprecipitated by incubating overnight with anti-TAK1 antibody and then incubated with protein A-Sepharose (PAS) for 4 h at 4°C. Precipitated proteins were separated by SDS-PAGE and immunoblotted to detect P-TAK1, TAK1, p-MKK4, MKK4, p-IKK α / β , IKK, and β -actin. Equivalent protein loading was verified by reprobing for the respective total antibodies and β -actin. Images are representative of 4 or more independent experiments. Values in bar graphs are means ± SE of at least 4 separate experiments performed in triplicate. ∗ P
Figure Legend Snippet: Torilin suppresses LPS-induced IKK and MAP3K complex activation and inhibits the TAK1 interaction with the components of MAPK and NF- κ B signaling pathways. RAW macrophage cells were incubated with torilin or vehicle for 30 min and then stimulated with LPS as indicated in the figures. (a) Lysates from torilin or vehicle treated-cells were immunoblotted to detect the activation status of IKK. (b) Immunoblots depicting the time dependent effects of torilin in TAK1, MKK4, and IKK phosphorylation. (c) The kinase activity of the TAK1 was determined after immunoprecipitates were resuspended in kinase buffer and assayed for kinase activity as described in Materials and Methods. (d) Cell lysates were immunoprecipitated by incubating overnight with anti-TAK1 antibody and then incubated with protein A-Sepharose (PAS) for 4 h at 4°C. Precipitated proteins were separated by SDS-PAGE and immunoblotted to detect P-TAK1, TAK1, p-MKK4, MKK4, p-IKK α / β , IKK, and β -actin. Equivalent protein loading was verified by reprobing for the respective total antibodies and β -actin. Images are representative of 4 or more independent experiments. Values in bar graphs are means ± SE of at least 4 separate experiments performed in triplicate. ∗ P

Techniques Used: Activation Assay, Incubation, Western Blot, Activity Assay, Immunoprecipitation, SDS Page

11) Product Images from "Shikonin suppresses ERK 1/2 phosphorylation during the early stages of adipocyte differentiation in 3T3-L1 cells"

Article Title: Shikonin suppresses ERK 1/2 phosphorylation during the early stages of adipocyte differentiation in 3T3-L1 cells

Journal: BMC Complementary and Alternative Medicine

doi: 10.1186/1472-6882-13-207

Effect of shikonin on adipogenic transcription factor protein levels and gene expression in 3T3-L1 adipocytes. (A) Western blot analysis was performed with antibodies against PPARγ, C/EBPα, aP2, and β-actin. (B) The expression of PPARγ, C/EBPα, and aP2mRNAs from 3T3-L1 adipocytes was measured by qRT-PCR.
Figure Legend Snippet: Effect of shikonin on adipogenic transcription factor protein levels and gene expression in 3T3-L1 adipocytes. (A) Western blot analysis was performed with antibodies against PPARγ, C/EBPα, aP2, and β-actin. (B) The expression of PPARγ, C/EBPα, and aP2mRNAs from 3T3-L1 adipocytes was measured by qRT-PCR.

Techniques Used: Expressing, Western Blot, Quantitative RT-PCR

Effect of shikonin on the lipid accumulation of 3T3-L1 adipocytes. (A) Cell viability was determined by an MTT assay. (B) The lipid content was assessed by Oil Red O staining. (C) The absorbance of Oil Red O dissolved in isoprophanol was spectrophotometrically determined at 500 nm.
Figure Legend Snippet: Effect of shikonin on the lipid accumulation of 3T3-L1 adipocytes. (A) Cell viability was determined by an MTT assay. (B) The lipid content was assessed by Oil Red O staining. (C) The absorbance of Oil Red O dissolved in isoprophanol was spectrophotometrically determined at 500 nm.

Techniques Used: MTT Assay, Staining

Inhibitory effect of shikonin on ERK 1/2 phosphorylation in 3T3-L1 adipocytes. (A) Protein level and (B) mRNA expression were determined with ERK 1/2. (C) Time-course of ERK expression as measured by qRT-PCR.
Figure Legend Snippet: Inhibitory effect of shikonin on ERK 1/2 phosphorylation in 3T3-L1 adipocytes. (A) Protein level and (B) mRNA expression were determined with ERK 1/2. (C) Time-course of ERK expression as measured by qRT-PCR.

Techniques Used: Expressing, Quantitative RT-PCR

12) Product Images from "dsRNA silencing of an R2R3-MYB transcription factor affects flower cell shape in a Dendrobium hybrid"

Article Title: dsRNA silencing of an R2R3-MYB transcription factor affects flower cell shape in a Dendrobium hybrid

Journal: BMC Plant Biology

doi: 10.1186/s12870-015-0577-3

Relative expression levels of the DhMYB1 cDNA in different floral tissues of floral buds. a Representative semi-quantitative RT-PCR results for DhMYB1 and β-actin cDNA in sepals, petals and labella of stage 5 to 7 floral buds. Lane 1: stage 5 sepals; lane 2: stage 5 petals; lane 3: stage 5 labella; lane 4: stage 6 sepals; lane 5: stage 6 petals; lane 6: stage 6 labella; lane 7: stage 7 sepals; lane 8: stage 7 petals; lane 9: stage 7 labella; lane 10: negative control. b Relative intensity of PCR bands for DhMYB1 normalized to the constitutive β-actin gene. Each bar represents the mean for three biological replicates with error bars indicating the SE. Letters indicate significant differences using one way ANOVA followed by Tukey HSD multi-comparison analysis test (P
Figure Legend Snippet: Relative expression levels of the DhMYB1 cDNA in different floral tissues of floral buds. a Representative semi-quantitative RT-PCR results for DhMYB1 and β-actin cDNA in sepals, petals and labella of stage 5 to 7 floral buds. Lane 1: stage 5 sepals; lane 2: stage 5 petals; lane 3: stage 5 labella; lane 4: stage 6 sepals; lane 5: stage 6 petals; lane 6: stage 6 labella; lane 7: stage 7 sepals; lane 8: stage 7 petals; lane 9: stage 7 labella; lane 10: negative control. b Relative intensity of PCR bands for DhMYB1 normalized to the constitutive β-actin gene. Each bar represents the mean for three biological replicates with error bars indicating the SE. Letters indicate significant differences using one way ANOVA followed by Tukey HSD multi-comparison analysis test (P

Techniques Used: Expressing, Quantitative RT-PCR, Negative Control, Polymerase Chain Reaction

13) Product Images from "Molecular cloning, purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein"

Article Title: Molecular cloning, purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein

Journal: Journal of Veterinary Science

doi: 10.4142/jvs.2016.17.1.119

Amplification of Brucella ( B. ) abortus mdh gene. PCR products on 1.5% (w/v) agarose gel. Lane 1, DNA marker; Lane 2, single expected band of mdh gene (963 bp, arrow).
Figure Legend Snippet: Amplification of Brucella ( B. ) abortus mdh gene. PCR products on 1.5% (w/v) agarose gel. Lane 1, DNA marker; Lane 2, single expected band of mdh gene (963 bp, arrow).

Techniques Used: Amplification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Marker

14) Product Images from "Torilin Inhibits Inflammation by Limiting TAK1-Mediated MAP Kinase and NF-κB Activation"

Article Title: Torilin Inhibits Inflammation by Limiting TAK1-Mediated MAP Kinase and NF-κB Activation

Journal: Mediators of Inflammation

doi: 10.1155/2017/7250968

Torilin restores I- κ B α  phosphorylation and degradation; thereby it inhibits p65-NF- κ B nuclear translocation. (a) RAW 264.7 cells were pretreated with the indicated concentrations of torilin for 30 min and incubated with LPS (100 ng/ml) for (15–60 min) time course and then assayed for the phosphorylation and degradation of I- κ B α  and nuclear translocation of p65 by western immunoblot analysis as described under Materials and Methods. (b and c) RAW macrophages were treated with LPS (100 ngmL −1 ) and 25  µ M torilin, respectively, for the indicated time course before cytoplasmic and nuclear protein fractions were subjected to western blot analyses for p65 (upper panel) and p50 (middle panel) of cytoplasmic (b) and nuclear (c) proteins, respectively.  β -Actin and ploy(ADP-ribose) polymerase (PARP) were used as a control for the cytoplasmic and nuclear protein loading, respectively. Images are representative of 3 or 4 independent experiments. Values in bar graphs are means ± SE of at least 4 independent experiments performed in triplicate. Significance was determined using Student's  t- test.  ∗ P
Figure Legend Snippet: Torilin restores I- κ B α phosphorylation and degradation; thereby it inhibits p65-NF- κ B nuclear translocation. (a) RAW 264.7 cells were pretreated with the indicated concentrations of torilin for 30 min and incubated with LPS (100 ng/ml) for (15–60 min) time course and then assayed for the phosphorylation and degradation of I- κ B α and nuclear translocation of p65 by western immunoblot analysis as described under Materials and Methods. (b and c) RAW macrophages were treated with LPS (100 ngmL −1 ) and 25  µ M torilin, respectively, for the indicated time course before cytoplasmic and nuclear protein fractions were subjected to western blot analyses for p65 (upper panel) and p50 (middle panel) of cytoplasmic (b) and nuclear (c) proteins, respectively. β -Actin and ploy(ADP-ribose) polymerase (PARP) were used as a control for the cytoplasmic and nuclear protein loading, respectively. Images are representative of 3 or 4 independent experiments. Values in bar graphs are means ± SE of at least 4 independent experiments performed in triplicate. Significance was determined using Student's t- test. ∗ P

Techniques Used: Translocation Assay, Incubation, Western Blot

Torilin arrests AP-1 and NF- κ B DNA binding and reporter gene expression in LPS-stimulated RAW 264.7 cells. RAW macrophages were treated with vehicle or torilin (as indicated) and stimulated with LPS for 45 min before nuclear protein was isolated. DNA binding was analyzed using specific  γ 32P-labeled oligonucleotide probes for NF- κ B (a) or AP-1 (b). Specificity was demonstrated by coincubation with a 25-fold excess of unlabeled specific probe of NF- κ B and AP-1 for competition. Cells were transiently transfected with NF- κ B (c) or AP-1 (d), plasmids treated with the indicated concentrations of torilin and LPS (100 ngmL −1 ) for 6 h and assayed for CAT expression using a CAT enzyme-linked immunosorbent assay kit. Each column shows the mean ± SEM of quadruplicate determinations. Luciferase activity was normalized to  β -galactosidase activity. Images are representative of 3 independent experiments.  ∗ P
Figure Legend Snippet: Torilin arrests AP-1 and NF- κ B DNA binding and reporter gene expression in LPS-stimulated RAW 264.7 cells. RAW macrophages were treated with vehicle or torilin (as indicated) and stimulated with LPS for 45 min before nuclear protein was isolated. DNA binding was analyzed using specific γ 32P-labeled oligonucleotide probes for NF- κ B (a) or AP-1 (b). Specificity was demonstrated by coincubation with a 25-fold excess of unlabeled specific probe of NF- κ B and AP-1 for competition. Cells were transiently transfected with NF- κ B (c) or AP-1 (d), plasmids treated with the indicated concentrations of torilin and LPS (100 ngmL −1 ) for 6 h and assayed for CAT expression using a CAT enzyme-linked immunosorbent assay kit. Each column shows the mean ± SEM of quadruplicate determinations. Luciferase activity was normalized to β -galactosidase activity. Images are representative of 3 independent experiments. ∗ P

Techniques Used: Binding Assay, Expressing, Isolation, Labeling, Transfection, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay

Torilin pretreatment reduces LPS-induced proinflammatory cytokine secretions into the culture media. RAW 264.7 cells were pretreated with torilin or vehicle for 30 min and stimulated with LPS for 24 h. TNF- α  (a), IL-1 β  (b), IL-6 (c), and GM-CSF (d) were determined using ELISA. Each bar graph represents mean ± (SE) for four independent experiments. Significance was determined using Student's  t -test.  ∗ P
Figure Legend Snippet: Torilin pretreatment reduces LPS-induced proinflammatory cytokine secretions into the culture media. RAW 264.7 cells were pretreated with torilin or vehicle for 30 min and stimulated with LPS for 24 h. TNF- α (a), IL-1 β (b), IL-6 (c), and GM-CSF (d) were determined using ELISA. Each bar graph represents mean ± (SE) for four independent experiments. Significance was determined using Student's t -test. ∗ P

Techniques Used: Enzyme-linked Immunosorbent Assay

Effect of torilin on mitogen activated protein kinase (MAPK) activation. LPS-stimulated RAW 264.7 macrophages were treated with vehicle or torilin (6.25–25  µ M) and stimulated with LPS for the indicated periods of time. Total protein was subjected to western blot analyses using total and phospho-anti-RK1/2, p38 MAPK , and JNK1/2 antibodies. Torilin suppressed phosphorylation of the three MAPKs (a and b). Cells were further pretreated with vehicle or PD98059 (30  µ M), ERK1/2 inhibitor, SB203580 (10  µ M), p38 MAPK  inhibitor, or SP600125 (10  µ M), JNK1/2 inhibitor, with torilin and stimulated with 100 ngmL −1  LPS as indicated in (c). Significance was determined using Student's  t- test versus the control group. Images are representative of 3 or 4 independent experiments. Values in bar graphs are means ± SE of at least 4 independent experiments performed in triplicate.  ∗ P
Figure Legend Snippet: Effect of torilin on mitogen activated protein kinase (MAPK) activation. LPS-stimulated RAW 264.7 macrophages were treated with vehicle or torilin (6.25–25  µ M) and stimulated with LPS for the indicated periods of time. Total protein was subjected to western blot analyses using total and phospho-anti-RK1/2, p38 MAPK , and JNK1/2 antibodies. Torilin suppressed phosphorylation of the three MAPKs (a and b). Cells were further pretreated with vehicle or PD98059 (30  µ M), ERK1/2 inhibitor, SB203580 (10  µ M), p38 MAPK inhibitor, or SP600125 (10  µ M), JNK1/2 inhibitor, with torilin and stimulated with 100 ngmL −1 LPS as indicated in (c). Significance was determined using Student's t- test versus the control group. Images are representative of 3 or 4 independent experiments. Values in bar graphs are means ± SE of at least 4 independent experiments performed in triplicate. ∗ P

Techniques Used: Activation Assay, Western Blot

15) Product Images from "Inhibition of Lipopolysaccharide-Induced Proinflammatory Responses by Buddleja officinalis Extract in BV-2 Microglial Cells via Negative Regulation of NF-κB and ERK1/2 Signaling"

Article Title: Inhibition of Lipopolysaccharide-Induced Proinflammatory Responses by Buddleja officinalis Extract in BV-2 Microglial Cells via Negative Regulation of NF-κB and ERK1/2 Signaling

Journal: Molecules

doi: 10.3390/molecules18089195

Effect of BOWE on LPS-induced mRNA expression levels of iNOS and COX-2 in BV-2 cells: For mRNA expression, BV-2 cells were pretreated with BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated, and mRNA levels of iNOS (A) and COX-2 (B) were then measured by RT-PCR. β actin expression was used as an internal control. The quantification of relative band intensities from three independent experiments were analyzed by densitometry and expressed as means ± SEM. * p
Figure Legend Snippet: Effect of BOWE on LPS-induced mRNA expression levels of iNOS and COX-2 in BV-2 cells: For mRNA expression, BV-2 cells were pretreated with BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated, and mRNA levels of iNOS (A) and COX-2 (B) were then measured by RT-PCR. β actin expression was used as an internal control. The quantification of relative band intensities from three independent experiments were analyzed by densitometry and expressed as means ± SEM. * p

Techniques Used: Expressing, Isolation, Reverse Transcription Polymerase Chain Reaction

Effect of BOWE on the mRNA expressions of IL-1β ( A ), IL-6 ( B ), and TNF-α ( C ): BV-2 cells were pretreated with or without the indicated concentrations of BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated and mRNA expressions of the aforementioned cytokines were measured by RT-PCR. β actin was used as a housekeeping gene. The quantification of relative band intensities from three independentexperiments were analyzed by densitometry and expressed as means ± SEM. * p
Figure Legend Snippet: Effect of BOWE on the mRNA expressions of IL-1β ( A ), IL-6 ( B ), and TNF-α ( C ): BV-2 cells were pretreated with or without the indicated concentrations of BOWE for 4 h and then stimulated with LPS (0.1 μg/mL) for 18 h. Total RNA was isolated and mRNA expressions of the aforementioned cytokines were measured by RT-PCR. β actin was used as a housekeeping gene. The quantification of relative band intensities from three independentexperiments were analyzed by densitometry and expressed as means ± SEM. * p

Techniques Used: Isolation, Reverse Transcription Polymerase Chain Reaction

16) Product Images from "Cell Growth of BG-1 Ovarian Cancer Cells was Promoted by 4-Tert-octylphenol and 4-Nonylphenol via Downregulation of TGF-? Receptor 2 and Upregulation of c-myc"

Article Title: Cell Growth of BG-1 Ovarian Cancer Cells was Promoted by 4-Tert-octylphenol and 4-Nonylphenol via Downregulation of TGF-? Receptor 2 and Upregulation of c-myc

Journal: Toxicological Research

doi: 10.5487/TR.2011.27.4.253

Altered expression levels of TGF-βR1 gene and TGF-βR2 gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated with E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). ( A ) Expression levels of TGF-βR1 by treatment with E2, OP or NP. ( B ) Expression levels of TGF-βR2 by treatment with E2, OP or NP. Expression level of TGF-βR1 and TGF-βR2 was detected by using semi-quantitative reverse-transcription PCR. Data represent the means ± S.D. of triplicate experiments. *, p
Figure Legend Snippet: Altered expression levels of TGF-βR1 gene and TGF-βR2 gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated with E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). ( A ) Expression levels of TGF-βR1 by treatment with E2, OP or NP. ( B ) Expression levels of TGF-βR2 by treatment with E2, OP or NP. Expression level of TGF-βR1 and TGF-βR2 was detected by using semi-quantitative reverse-transcription PCR. Data represent the means ± S.D. of triplicate experiments. *, p

Techniques Used: Expressing, Polymerase Chain Reaction

Altered expression levels of TGF-β1 gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated with E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). Expression level of TGF-β1 was detected by using semi-quantitative reverse-transcription PCR. PCR products were run on a 1.5% agarose gel, bands were scanned and the density of the bands on the gel was quantified using Gel Doc 2000 as described in Materials and Methods. Data represent the means ± S.D. of triplicate experiments. *, p
Figure Legend Snippet: Altered expression levels of TGF-β1 gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated with E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). Expression level of TGF-β1 was detected by using semi-quantitative reverse-transcription PCR. PCR products were run on a 1.5% agarose gel, bands were scanned and the density of the bands on the gel was quantified using Gel Doc 2000 as described in Materials and Methods. Data represent the means ± S.D. of triplicate experiments. *, p

Techniques Used: Expressing, Polymerase Chain Reaction, Agarose Gel Electrophoresis

Altered expression levels of c-myc gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). Expression level of c-myc was detected by using semi-quantitative reverse-transcription PCR. Data represent the means ± S.D. of triplicate experiments. *, p
Figure Legend Snippet: Altered expression levels of c-myc gene following treatments with E2, OP or NP. BG-1 cells were seeded in 6-well plates and treated E2 (10 -9 M), OP (10 -6 M) or NP (10 -6 M). Total RNAs were extracted in a time-dependent manner (0, 6, and 24 h). Expression level of c-myc was detected by using semi-quantitative reverse-transcription PCR. Data represent the means ± S.D. of triplicate experiments. *, p

Techniques Used: Expressing, Polymerase Chain Reaction

17) Product Images from "Molecular and immunological characterization of Hyalomma dromedarii and Hyalomma excavatum (Acari: Ixodidae) vectors of Q fever in camels"

Article Title: Molecular and immunological characterization of Hyalomma dromedarii and Hyalomma excavatum (Acari: Ixodidae) vectors of Q fever in camels

Journal: Veterinary World

doi: 10.14202/vetworld.2018.1109-1119

Agarose gel electrophoresis of polymerase chain reaction products obtained from H. dromedarii using IS30A spacer: (M) 1 kb DNA ladder, (N) negative control, lanes 7 and 8 represent positive tick samples and lanes 1-6 represent negative tick samples.
Figure Legend Snippet: Agarose gel electrophoresis of polymerase chain reaction products obtained from H. dromedarii using IS30A spacer: (M) 1 kb DNA ladder, (N) negative control, lanes 7 and 8 represent positive tick samples and lanes 1-6 represent negative tick samples.

Techniques Used: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Negative Control

Agarose gel electrophoresis of polymerase chain reaction products obtained from Hyalomma dromedarii and Hyalomma excavatum using two genes: (a) 16S ribosomal RNA gene reveals 440 bp, (b) cytochrome oxidase subunit-1 gene, reveals 850 bp, (M) 1 kb DNA ladder, (N) negative control.
Figure Legend Snippet: Agarose gel electrophoresis of polymerase chain reaction products obtained from Hyalomma dromedarii and Hyalomma excavatum using two genes: (a) 16S ribosomal RNA gene reveals 440 bp, (b) cytochrome oxidase subunit-1 gene, reveals 850 bp, (M) 1 kb DNA ladder, (N) negative control.

Techniques Used: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Negative Control

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Amplification:

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Synthesized:

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Isolation:

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Spectrophotometry:

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Real-time Polymerase Chain Reaction:

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Concentration Assay:

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Incubation:

Article Title: Co-treatment with therapeutic neural stem cells expressing carboxyl esterase and CPT-11 inhibit growth of primary and metastatic lung cancers in mice
Article Snippet: .. Briefly, 1 μg of RNA from HB1.F3.CE and A549 lung cancer cells was reverse transcribed with murine leukemia virus reverse transcriptase (MMLV-RT; iNtRON Biotechnology, Sungnam, Kyeonggido, Korea), 10 pM dNTP (Bioneer, Deajeon, Korea), 200 pM monomer random primer (TaKaRa Bio., Shiga, Japan), 5 × RT buffer (iNtRON Biotechnology) and RNase inhibitor (iNtRON Biotechnology). cDNA was then synthesized at 37°C for one hour, after which the reaction was stopped by incubation at 95°C for 5 min. .. The cDNA produced from the extracted total RNA of HB1.F3.CE and A549 lung cancer cells was amplified by PCR to confirm the expression of human or rabbit CE genes.

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Polymerase Chain Reaction:

Article Title: Effects of Genetically Engineered Stem Cells Expressing Cytosine Deaminase and Interferon-Beta or Carboxyl Esterase on the Growth of LNCaP Prostate Cancer Cells
Article Snippet: .. Concentration of the isolated total RNA was measured with a spectrophotometer (Optizen, Mecasys, Daejeon, Republic of Korea) and 1 μg of total RNA was used to synthesize cDNA using murine leukemia virus reverse transcriptase (M-MLV RT; iNtRON Biotechnology, Sungnam, Korea), 200 pM nonamer random primer (TaKaRa Co., Ltd., Osaka, Japan), 10 mM dNTPs (iNtRON Biotechnology), 4 μL 5× RT buffer (iNtRON Biotechnology), and 10 unit RNase inhibitor (iNtRON Biotechnology) at 37 °C for 1 h. The cDNA template was amplified by PCR in a mixture of 2.5 Unit Taq polymerase (iNtRON Biotechnology), 2 μL 10× PCR buffer (iNtRON Biotechnology), 10 mM dNTPs (iNtRON Biotechnology), and 200 pmol/L reverse and forward primers specific for CE, CD, IFN-β, and GAPDH (Bioneer Corporation, Daejoen, Republic of Korea) using a PTC-100 thermocycler (MJ Research Inc., Waltham, MA, USA). .. PCR was performed for 30 cycles of denaturation at 95 °C for 30 s, annealing at 58 °C for 30 s, and extension at 72 °C for 30 s. The primer sequences and predicted product sizes are shown in .

Article Title: Immunomodulatory Efficacy of Standardized Annona muricata (Graviola) Leaf Extract via Activation of Mitogen-Activated Protein Kinase Pathways in RAW 264.7 Macrophages
Article Snippet: .. RNA Isolation and Real-Time RT-PCR RAW 264.7 cells were treated with 0, 50, 100, and 200 μ g/mL GE for 8 h after 8 × 105 cells/well were seeded into a 6-well plate and incubated for 24 h. Total RNA was isolated from RAW 264.7 cells using RNA Extraction Kit (Intron Biotechnology Inc., Korea) according to the manufacturer's instructions. cDNA was synthesized in a PCR Thermal Cycler (TaKaRa, Japan), using the iScript cDNA synthesis Kit (Bio-Rad Laboratories, Hercules, CA, USA). ..

Article Title: Molecular and immunological characterization of Hyalomma dromedarii and Hyalomma excavatum (Acari: Ixodidae) vectors of Q fever in camels
Article Snippet: .. Each PCR mixture contained 25-50 ng/µl genomic DNA, 10 pM/µl of each primer, 12.5 µl of 2× PCR Master Mix solution (i-Taq) PCR master mix (1× buffer, 2.5 mM dNTP, 1× gel loading buffer, and 4 mM MgCl2 ; iNtRon Biotechnology, Korea), and 9 µl nuclease-free water (Qiagen) to complete the total volume of the reaction. .. All amplifications were performed in BIO-RAD Thermal Cycler (BIO-RAD, Singapore) utilizing the following cycling profile, for the two DNA markers one cycle at 94°C for 5 min (initial denaturation).

Quantitative RT-PCR:

Article Title: Immunomodulatory Efficacy of Standardized Annona muricata (Graviola) Leaf Extract via Activation of Mitogen-Activated Protein Kinase Pathways in RAW 264.7 Macrophages
Article Snippet: .. RNA Isolation and Real-Time RT-PCR RAW 264.7 cells were treated with 0, 50, 100, and 200 μ g/mL GE for 8 h after 8 × 105 cells/well were seeded into a 6-well plate and incubated for 24 h. Total RNA was isolated from RAW 264.7 cells using RNA Extraction Kit (Intron Biotechnology Inc., Korea) according to the manufacturer's instructions. cDNA was synthesized in a PCR Thermal Cycler (TaKaRa, Japan), using the iScript cDNA synthesis Kit (Bio-Rad Laboratories, Hercules, CA, USA). ..

Article Title: Gene Expression Profiling during Conidiation in the Rice Blast Pathogen Magnaporthe oryzae
Article Snippet: .. Total RNA was isolated from non-condiating mycelia and conidiating mycelia using the Easy-Spin total RNA extraction kit (Intron Biotechnology, Seongnam, Korea); total RNA was treated with DNase prior to use in microarray or qRT-PCR. .. For harvesting non-conidiating mycelia, agar plugs (5 mm in diameter) were obtained from the actively growing edge of oatmeal agar (OMA) plates, inoculated into liquid complete medium, and incubated at 28°C on a 250 rpm shaker for 4 days.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Isolation and Characterization of a Theta Glutathione S-transferase Gene from Panax ginseng Meyer
Article Snippet: .. For RT-PCR, 200 ng of total RNA was used as a template for reverse transcription using oligo (dT)15 primer (0.2 mM) and AMV Reverse Transcriptase (10 U/μL; Intron Biotechnology Inc., Seongnam, Korea) according to the manufacturer’s instructions. .. The PCR reaction was then performed using a 1 μL aliquot of the first strand cDNA in a final volume of 20 μL containing 5 pmol of specific primers for coding region of PgGST (forward, 5’-CGT TCT CAT CTT CTG CAA GGT C-3’; reverse, 5’-GCT GAG ATC TGC TAT GGA TGG T-3’).

Microarray:

Article Title: Gene Expression Profiling during Conidiation in the Rice Blast Pathogen Magnaporthe oryzae
Article Snippet: .. Total RNA was isolated from non-condiating mycelia and conidiating mycelia using the Easy-Spin total RNA extraction kit (Intron Biotechnology, Seongnam, Korea); total RNA was treated with DNase prior to use in microarray or qRT-PCR. .. For harvesting non-conidiating mycelia, agar plugs (5 mm in diameter) were obtained from the actively growing edge of oatmeal agar (OMA) plates, inoculated into liquid complete medium, and incubated at 28°C on a 250 rpm shaker for 4 days.

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    iNtRON Biotechnology pcr mixture
    Agarose gel electrophoresis of polymerase chain reaction products obtained from Hyalomma dromedarii and Hyalomma excavatum using two genes: (a) 16S ribosomal RNA gene reveals 440 bp, (b) cytochrome oxidase subunit-1 gene, reveals 850 bp, (M) 1 kb <t>DNA</t> ladder, (N) negative control.
    Pcr Mixture, supplied by iNtRON Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr mixture/product/iNtRON Biotechnology
    Average 93 stars, based on 23 article reviews
    Price from $9.99 to $1999.99
    pcr mixture - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    89
    iNtRON Biotechnology pcr master mix solution
    Amplification of Brucella ( B. ) abortus mdh gene. <t>PCR</t> products on 1.5% (w/v) agarose gel. Lane 1, <t>DNA</t> marker; Lane 2, single expected band of mdh gene (963 bp, arrow).
    Pcr Master Mix Solution, supplied by iNtRON Biotechnology, used in various techniques. Bioz Stars score: 89/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr master mix solution/product/iNtRON Biotechnology
    Average 89 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    pcr master mix solution - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    91
    iNtRON Biotechnology total pcr reaction mixture
    Identification of strains MJM60319, MJM60298, and MJM60399. a Morphology of strains MJM60319, MJM60298, and MJM60399 observed by scanning electron microscopy. Scale bar indicates 1 µm, b phylogenetic tree constructed based on 16S rDNA sequence of the strain. The strains MJM60319, MJM60298, and MJM60399 possesses 99% similarity with L. plantarum . c GTG5 <t>PCR</t> fingerprinting analysis of LAB strains. 1, MJM60319; 2, MJM60298; 3, MJM60399; 4, L. plantarum ssp. plantarum KACC 11451; 5, L. paraplantarum KACC 12373; 6, L. pentosus KACC 12428; M, Marker: 100–10,000 bp GeneRulerTM <t>DNA</t> ladder mix (Fermentas)
    Total Pcr Reaction Mixture, supplied by iNtRON Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/total pcr reaction mixture/product/iNtRON Biotechnology
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    total pcr reaction mixture - by Bioz Stars, 2020-09
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    Agarose gel electrophoresis of polymerase chain reaction products obtained from Hyalomma dromedarii and Hyalomma excavatum using two genes: (a) 16S ribosomal RNA gene reveals 440 bp, (b) cytochrome oxidase subunit-1 gene, reveals 850 bp, (M) 1 kb DNA ladder, (N) negative control.

    Journal: Veterinary World

    Article Title: Molecular and immunological characterization of Hyalomma dromedarii and Hyalomma excavatum (Acari: Ixodidae) vectors of Q fever in camels

    doi: 10.14202/vetworld.2018.1109-1119

    Figure Lengend Snippet: Agarose gel electrophoresis of polymerase chain reaction products obtained from Hyalomma dromedarii and Hyalomma excavatum using two genes: (a) 16S ribosomal RNA gene reveals 440 bp, (b) cytochrome oxidase subunit-1 gene, reveals 850 bp, (M) 1 kb DNA ladder, (N) negative control.

    Article Snippet: Each PCR mixture contained 25-50 ng/µl genomic DNA, 10 pM/µl of each primer, 12.5 µl of 2× PCR Master Mix solution (i-Taq) PCR master mix (1× buffer, 2.5 mM dNTP, 1× gel loading buffer, and 4 mM MgCl2 ; iNtRon Biotechnology, Korea), and 9 µl nuclease-free water (Qiagen) to complete the total volume of the reaction.

    Techniques: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Negative Control

    Agarose gel electrophoresis of polymerase chain reaction products obtained from H. dromedarii using IS30A spacer: (M) 1 kb DNA ladder, (N) negative control, lanes 7 and 8 represent positive tick samples and lanes 1-6 represent negative tick samples.

    Journal: Veterinary World

    Article Title: Molecular and immunological characterization of Hyalomma dromedarii and Hyalomma excavatum (Acari: Ixodidae) vectors of Q fever in camels

    doi: 10.14202/vetworld.2018.1109-1119

    Figure Lengend Snippet: Agarose gel electrophoresis of polymerase chain reaction products obtained from H. dromedarii using IS30A spacer: (M) 1 kb DNA ladder, (N) negative control, lanes 7 and 8 represent positive tick samples and lanes 1-6 represent negative tick samples.

    Article Snippet: Each PCR mixture contained 25-50 ng/µl genomic DNA, 10 pM/µl of each primer, 12.5 µl of 2× PCR Master Mix solution (i-Taq) PCR master mix (1× buffer, 2.5 mM dNTP, 1× gel loading buffer, and 4 mM MgCl2 ; iNtRon Biotechnology, Korea), and 9 µl nuclease-free water (Qiagen) to complete the total volume of the reaction.

    Techniques: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Negative Control

    Relative expression levels of the DhMYB1 cDNA in different floral tissues of floral buds. a Representative semi-quantitative RT-PCR results for DhMYB1 and β-actin cDNA in sepals, petals and labella of stage 5 to 7 floral buds. Lane 1: stage 5 sepals; lane 2: stage 5 petals; lane 3: stage 5 labella; lane 4: stage 6 sepals; lane 5: stage 6 petals; lane 6: stage 6 labella; lane 7: stage 7 sepals; lane 8: stage 7 petals; lane 9: stage 7 labella; lane 10: negative control. b Relative intensity of PCR bands for DhMYB1 normalized to the constitutive β-actin gene. Each bar represents the mean for three biological replicates with error bars indicating the SE. Letters indicate significant differences using one way ANOVA followed by Tukey HSD multi-comparison analysis test (P

    Journal: BMC Plant Biology

    Article Title: dsRNA silencing of an R2R3-MYB transcription factor affects flower cell shape in a Dendrobium hybrid

    doi: 10.1186/s12870-015-0577-3

    Figure Lengend Snippet: Relative expression levels of the DhMYB1 cDNA in different floral tissues of floral buds. a Representative semi-quantitative RT-PCR results for DhMYB1 and β-actin cDNA in sepals, petals and labella of stage 5 to 7 floral buds. Lane 1: stage 5 sepals; lane 2: stage 5 petals; lane 3: stage 5 labella; lane 4: stage 6 sepals; lane 5: stage 6 petals; lane 6: stage 6 labella; lane 7: stage 7 sepals; lane 8: stage 7 petals; lane 9: stage 7 labella; lane 10: negative control. b Relative intensity of PCR bands for DhMYB1 normalized to the constitutive β-actin gene. Each bar represents the mean for three biological replicates with error bars indicating the SE. Letters indicate significant differences using one way ANOVA followed by Tukey HSD multi-comparison analysis test (P

    Article Snippet: Semi-quantitative PCR was performed using a 20 μL PCR mixture containing 2 μL of cDNA, 5 pmoles of each primer, 1X PCR buffer, 0.25 mM of each dNTP and 1U of i-Taq DNA polymerase (iNtRON Biotechnology, Inc., Korea).

    Techniques: Expressing, Quantitative RT-PCR, Negative Control, Polymerase Chain Reaction

    Amplification of Brucella ( B. ) abortus mdh gene. PCR products on 1.5% (w/v) agarose gel. Lane 1, DNA marker; Lane 2, single expected band of mdh gene (963 bp, arrow).

    Journal: Journal of Veterinary Science

    Article Title: Molecular cloning, purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein

    doi: 10.4142/jvs.2016.17.1.119

    Figure Lengend Snippet: Amplification of Brucella ( B. ) abortus mdh gene. PCR products on 1.5% (w/v) agarose gel. Lane 1, DNA marker; Lane 2, single expected band of mdh gene (963 bp, arrow).

    Article Snippet: The PCR mixture included 4 µL of bacterial genomic DNA, 25 µL of 2× PCR Master Mix Solution (iNtRON Biotechnology, Korea) and 10 picomoles of forward and reverse primers diluted to a final volume of 50 µL.

    Techniques: Amplification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Marker

    Identification of strains MJM60319, MJM60298, and MJM60399. a Morphology of strains MJM60319, MJM60298, and MJM60399 observed by scanning electron microscopy. Scale bar indicates 1 µm, b phylogenetic tree constructed based on 16S rDNA sequence of the strain. The strains MJM60319, MJM60298, and MJM60399 possesses 99% similarity with L. plantarum . c GTG5 PCR fingerprinting analysis of LAB strains. 1, MJM60319; 2, MJM60298; 3, MJM60399; 4, L. plantarum ssp. plantarum KACC 11451; 5, L. paraplantarum KACC 12373; 6, L. pentosus KACC 12428; M, Marker: 100–10,000 bp GeneRulerTM DNA ladder mix (Fermentas)

    Journal: Indian Journal of Microbiology

    Article Title: In Vitro Characterization of Lactobacillus plantarum Strains with Inhibitory Activity on Enteropathogens for Use as Potential Animal Probiotics

    doi: 10.1007/s12088-017-0646-4

    Figure Lengend Snippet: Identification of strains MJM60319, MJM60298, and MJM60399. a Morphology of strains MJM60319, MJM60298, and MJM60399 observed by scanning electron microscopy. Scale bar indicates 1 µm, b phylogenetic tree constructed based on 16S rDNA sequence of the strain. The strains MJM60319, MJM60298, and MJM60399 possesses 99% similarity with L. plantarum . c GTG5 PCR fingerprinting analysis of LAB strains. 1, MJM60319; 2, MJM60298; 3, MJM60399; 4, L. plantarum ssp. plantarum KACC 11451; 5, L. paraplantarum KACC 12373; 6, L. pentosus KACC 12428; M, Marker: 100–10,000 bp GeneRulerTM DNA ladder mix (Fermentas)

    Article Snippet: A 20 μl total PCR reaction mixture consisted of 1 μl genomic DNA (50 ng), primer 1 μl (100 pmol), and 18 μl sterile deionized water added to maxime i-Taq PCR premix tubes (iNtRON Biotechnology, INC, Gyeonggi-do, Korea).

    Techniques: Electron Microscopy, Construct, Sequencing, Polymerase Chain Reaction, Marker