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Kapa Biosystems kapa real time amplification kit
Kapa Real Time Amplification Kit, supplied by Kapa Biosystems, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/kapa real time amplification kit/product/Kapa Biosystems
Average 91 stars, based on 5 article reviews
Price from $9.99 to $1999.99
kapa real time amplification kit - by Bioz Stars, 2020-07
91/100 stars

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Polymerase Chain Reaction:

Article Title: Multifunctional role of the transcription factor Blimp1 in coordinating plasma cell differentiation
Article Snippet: .. For strand-specific RNA-sequencing, the uridines present in one cDNA strand were digested with uracil-N-glycosylase (New England Biolabs) as described followed by PCR amplification with the KAPA Real Time Amplification kit (KAPA Biosystems). .. Completed libraries were quantified with the Agilent Bioanalyzer dsDNA 1000 assay kit and Agilent QPCR NGS Library Quantification kit.

Real-time Polymerase Chain Reaction:

Article Title: Whole genome screen reveals a novel relationship between Wolbachia levels and Drosophila host translation
Article Snippet: .. After adaptor ligation, the libraries were amplified by qPCR using the KAPA Real-time amplification kit (KAPA Biosystems). .. Finally, libraries were purified using Agencourt AMPure XP beads (Beckman Coulter) as described in the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England BioLabs, E7420L) protocol.

Amplification:

Article Title: Targeted genome fragmentation with CRISPR/Cas9 enables fast and efficient enrichment of small genomic regions and ultra-accurate sequencing with low DNA input (CRISPR-DS)
Article Snippet: .. The ligated DNA was amplified using KAPA Real-Time Amplification kit with fluorescent standards (KAPA Biosystems). .. Fifty-microliter reactions were prepared including KAPA HiFi HotStart Real-time PCR Master Mix, 23 µL of previously ligated and purified DNA, and DS primers MWS13, 5′-AATGATACGGCGACCACCGAG-3′, and MWS20, 5′-GTGACTGGAGTTCAGACGTGTGC-3′ ( ; ) at a final concentration of 2 µM.

Article Title: Multifunctional role of the transcription factor Blimp1 in coordinating plasma cell differentiation
Article Snippet: .. For strand-specific RNA-sequencing, the uridines present in one cDNA strand were digested with uracil-N-glycosylase (New England Biolabs) as described followed by PCR amplification with the KAPA Real Time Amplification kit (KAPA Biosystems). .. Completed libraries were quantified with the Agilent Bioanalyzer dsDNA 1000 assay kit and Agilent QPCR NGS Library Quantification kit.

Article Title: Whole genome screen reveals a novel relationship between Wolbachia levels and Drosophila host translation
Article Snippet: .. After adaptor ligation, the libraries were amplified by qPCR using the KAPA Real-time amplification kit (KAPA Biosystems). .. Finally, libraries were purified using Agencourt AMPure XP beads (Beckman Coulter) as described in the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England BioLabs, E7420L) protocol.

RNA Sequencing Assay:

Article Title: Multifunctional role of the transcription factor Blimp1 in coordinating plasma cell differentiation
Article Snippet: .. For strand-specific RNA-sequencing, the uridines present in one cDNA strand were digested with uracil-N-glycosylase (New England Biolabs) as described followed by PCR amplification with the KAPA Real Time Amplification kit (KAPA Biosystems). .. Completed libraries were quantified with the Agilent Bioanalyzer dsDNA 1000 assay kit and Agilent QPCR NGS Library Quantification kit.

Ligation:

Article Title: Whole genome screen reveals a novel relationship between Wolbachia levels and Drosophila host translation
Article Snippet: .. After adaptor ligation, the libraries were amplified by qPCR using the KAPA Real-time amplification kit (KAPA Biosystems). .. Finally, libraries were purified using Agencourt AMPure XP beads (Beckman Coulter) as described in the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England BioLabs, E7420L) protocol.

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    Kapa Biosystems qpcr kit
    ME1 knockdown mimicked the inhibitory effect of miR-612 overexpression in bladder cancer cells. (A) <t>mRNA</t> and (B) protein levels of ME1 in T24 cells following transfection with si-ME1 or si-NC were measured using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. GAPDH and β-actin were used as internal controls, respectively. (C) Cell proliferation, (D) colony formation, (E) Transwell tumor cell migration assay and (F) Transwell tumor cell invasion assays in bladder cancer T24 cells transfected with si-ME1 or si-NC. (G) Protein levels of EMT-associated markers in T24 cells transfected with si-ME1 or si-NC were evaluated by western blot analysis. * P
    Qpcr Kit, supplied by Kapa Biosystems, used in various techniques. Bioz Stars score: 94/100, based on 170 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qpcr kit/product/Kapa Biosystems
    Average 94 stars, based on 170 article reviews
    Price from $9.99 to $1999.99
    qpcr kit - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    94
    Roche pcr universal kit
    Real-time <t>PCR</t> and Western blotting analyses of CLIC2, CLIC3 and CLIC4 in ESCCs and paired NTs. (A-C): CLIC2 (A), CLIC3 (B) and CLIC4 (C) <t>mRNA</t> assays by qPCR between ESCCs and paired NTs. (D-F): Representative CLIC2 (D), CLIC3 (E) and CLIC4 (F) protein assays by Western blotting in ESCCs and paired NTs, GAPDH served as loading control. ( G-I): Statistical analyses of CLIC2 (G), CLIC3 (H) and CLIC4 (I) protein levels between ESCCs and paired NTs.
    Pcr Universal Kit, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr universal kit/product/Roche
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcr universal kit - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

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    ME1 knockdown mimicked the inhibitory effect of miR-612 overexpression in bladder cancer cells. (A) mRNA and (B) protein levels of ME1 in T24 cells following transfection with si-ME1 or si-NC were measured using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. GAPDH and β-actin were used as internal controls, respectively. (C) Cell proliferation, (D) colony formation, (E) Transwell tumor cell migration assay and (F) Transwell tumor cell invasion assays in bladder cancer T24 cells transfected with si-ME1 or si-NC. (G) Protein levels of EMT-associated markers in T24 cells transfected with si-ME1 or si-NC were evaluated by western blot analysis. * P

    Journal: International Journal of Oncology

    Article Title: Tumor-suppressing effects of microRNA-612 in bladder cancer cells by targeting malic enzyme 1 expression

    doi: 10.3892/ijo.2018.4342

    Figure Lengend Snippet: ME1 knockdown mimicked the inhibitory effect of miR-612 overexpression in bladder cancer cells. (A) mRNA and (B) protein levels of ME1 in T24 cells following transfection with si-ME1 or si-NC were measured using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. GAPDH and β-actin were used as internal controls, respectively. (C) Cell proliferation, (D) colony formation, (E) Transwell tumor cell migration assay and (F) Transwell tumor cell invasion assays in bladder cancer T24 cells transfected with si-ME1 or si-NC. (G) Protein levels of EMT-associated markers in T24 cells transfected with si-ME1 or si-NC were evaluated by western blot analysis. * P

    Article Snippet: The amplification procedure was as follows: 5 min at 95°C, followed by 40 cycles at 95°C for 30 sec and 65°C for 45 sec. For detection of the ME1 mRNA level, cDNA was synthesized using the PrimeScript RT Reagent kit (Takara Bio, Inc., Tokyo, Japan), according to the manufacturer's protocol. qPCR for mRNA detection was also performed using the KAPA SYBR FAST qPCR kit (Kapa Biosystems, Inc.), The primers used were as follows: ME1 forward, 5′-GCAGGTCTCCTTGCAGCTCT-3′ and reverse, 5′-TCCAAGGCCATCACA ATCAG-3′; GAPDH forward, 5′-ATGTCGTGGAGTCTACTGGC-3′ and reverse, 5′-TGACCTTGCCCACAGCCTTG-3′.

    Techniques: Over Expression, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Cell Migration Assay

    ME1 overexpression partially rescues the inhibitory effects of miR-612 expression on bladder cancer cells. (A) mRNA and (B) protein levels of ME1 in T24 cells co-transfected with miR-612 mimic or miR-NC, and with or without ME1 cDNA were measured using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. (C) Cell proliferation, (D) colony formation, (E) Transwell tumor cell migration and (F) Transwell tumor cell invasion assays in T24 cells co-transfected with miR-612 mimic or miR-NC, and with or without ME1 cDNA. * P

    Journal: International Journal of Oncology

    Article Title: Tumor-suppressing effects of microRNA-612 in bladder cancer cells by targeting malic enzyme 1 expression

    doi: 10.3892/ijo.2018.4342

    Figure Lengend Snippet: ME1 overexpression partially rescues the inhibitory effects of miR-612 expression on bladder cancer cells. (A) mRNA and (B) protein levels of ME1 in T24 cells co-transfected with miR-612 mimic or miR-NC, and with or without ME1 cDNA were measured using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. (C) Cell proliferation, (D) colony formation, (E) Transwell tumor cell migration and (F) Transwell tumor cell invasion assays in T24 cells co-transfected with miR-612 mimic or miR-NC, and with or without ME1 cDNA. * P

    Article Snippet: The amplification procedure was as follows: 5 min at 95°C, followed by 40 cycles at 95°C for 30 sec and 65°C for 45 sec. For detection of the ME1 mRNA level, cDNA was synthesized using the PrimeScript RT Reagent kit (Takara Bio, Inc., Tokyo, Japan), according to the manufacturer's protocol. qPCR for mRNA detection was also performed using the KAPA SYBR FAST qPCR kit (Kapa Biosystems, Inc.), The primers used were as follows: ME1 forward, 5′-GCAGGTCTCCTTGCAGCTCT-3′ and reverse, 5′-TCCAAGGCCATCACA ATCAG-3′; GAPDH forward, 5′-ATGTCGTGGAGTCTACTGGC-3′ and reverse, 5′-TGACCTTGCCCACAGCCTTG-3′.

    Techniques: Over Expression, Expressing, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Migration

    ME1 is a direct target of miR-612. (A) Bioinformatic analysis indicated that ME1 is the target of miR-612. The presumable miR-612-binding sites in the 3′-UTR of ME1 cDNA and the mutant binding sites are shown. (B) The relative luciferase activity was determined in T24 cells following co-transfection with a luciferase reporter plasmid (WT or MUT 3′-UTR ME1 cDNA), and miR-612 mimic or miR-NC. (C) mRNA and (D) protein levels of ME1 in T24 cells following transfection with miR-612 mimic or miR-NC were analyzed using RT-qPCR and western blot analysis, respectively. GAPDH and β-actin served as the internal controls, respectively. (E) The levels of ME1 mRNA were measured using RT-qPCR in 46 paired bladder cancer and ANT specimens. GAPDH was used as an internal control. (F) The correlation between ME1 and miR-612 expression in bladder cancer tissue samples (n=46) was analyzed using Spearman's correlation test. ** P

    Journal: International Journal of Oncology

    Article Title: Tumor-suppressing effects of microRNA-612 in bladder cancer cells by targeting malic enzyme 1 expression

    doi: 10.3892/ijo.2018.4342

    Figure Lengend Snippet: ME1 is a direct target of miR-612. (A) Bioinformatic analysis indicated that ME1 is the target of miR-612. The presumable miR-612-binding sites in the 3′-UTR of ME1 cDNA and the mutant binding sites are shown. (B) The relative luciferase activity was determined in T24 cells following co-transfection with a luciferase reporter plasmid (WT or MUT 3′-UTR ME1 cDNA), and miR-612 mimic or miR-NC. (C) mRNA and (D) protein levels of ME1 in T24 cells following transfection with miR-612 mimic or miR-NC were analyzed using RT-qPCR and western blot analysis, respectively. GAPDH and β-actin served as the internal controls, respectively. (E) The levels of ME1 mRNA were measured using RT-qPCR in 46 paired bladder cancer and ANT specimens. GAPDH was used as an internal control. (F) The correlation between ME1 and miR-612 expression in bladder cancer tissue samples (n=46) was analyzed using Spearman's correlation test. ** P

    Article Snippet: The amplification procedure was as follows: 5 min at 95°C, followed by 40 cycles at 95°C for 30 sec and 65°C for 45 sec. For detection of the ME1 mRNA level, cDNA was synthesized using the PrimeScript RT Reagent kit (Takara Bio, Inc., Tokyo, Japan), according to the manufacturer's protocol. qPCR for mRNA detection was also performed using the KAPA SYBR FAST qPCR kit (Kapa Biosystems, Inc.), The primers used were as follows: ME1 forward, 5′-GCAGGTCTCCTTGCAGCTCT-3′ and reverse, 5′-TCCAAGGCCATCACA ATCAG-3′; GAPDH forward, 5′-ATGTCGTGGAGTCTACTGGC-3′ and reverse, 5′-TGACCTTGCCCACAGCCTTG-3′.

    Techniques: Binding Assay, Mutagenesis, Luciferase, Activity Assay, Cotransfection, Plasmid Preparation, Transfection, Quantitative RT-PCR, Western Blot, Expressing

    RSV effects on IL-17A, IL-19 and IL-23p19 gene expression. Quantitative PCR of IL-17A, IL-19 and IL-23p19 gene expression was determined to quantify effects of RSV on IL-17A, IL-19 and IL-23p19 gene expression. The mRNA levels of IL-17A, IL-19 and IL-23p19 were quantified using MYO18B as reference gene. Clamped bar with * above indicates the pair of column means are significantly different (p

    Journal: PLoS ONE

    Article Title: Resveratrol Ameliorates Imiquimod-Induced Psoriasis-Like Skin Inflammation in Mice

    doi: 10.1371/journal.pone.0126599

    Figure Lengend Snippet: RSV effects on IL-17A, IL-19 and IL-23p19 gene expression. Quantitative PCR of IL-17A, IL-19 and IL-23p19 gene expression was determined to quantify effects of RSV on IL-17A, IL-19 and IL-23p19 gene expression. The mRNA levels of IL-17A, IL-19 and IL-23p19 were quantified using MYO18B as reference gene. Clamped bar with * above indicates the pair of column means are significantly different (p

    Article Snippet: The PCR reactions were performed in duplicates using the KAPA SYBR FAST qPCR kit (Kapa Biosystems, Inc., Woburn, MA) in a LightCycler 480 (Roche Applied Science) using the following protocol: One step at 95°C for 3 min, then 95°C for 10 s, 60°C for 20 s, and 72°C for 10 s. The increase in fluorescence was measured in real time during the extension step and a final melt curve analysis was performed to verify the specificity of the amplification.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Quantitative PCR of microarray genes. Selected qPCR of genes that were 1.5 fold or more changed by RSV treatment in the microarray (RSV treated compared with the IMQ group). The mRNA levels were quantified using MYO18B as reference gene. (a) Phosphoenolpyruvate Carboxykinase 1 (PCK1). (b) Tripartite Motif Containing 63, E3 Ubiquitin Protein Ligase (TRIM63). (c) Protein Phosphatase 1, Regulatory (inhibitor) Subunit 3C (PPP1R3C). Columns in (a-c) are group means ±SEM (n = 8, n = 10, n = 10 for controls, IMQ, IMQ-RSV respectively). Clamped bar with * above indicates the pair of column means are significantly different (p

    Journal: PLoS ONE

    Article Title: Resveratrol Ameliorates Imiquimod-Induced Psoriasis-Like Skin Inflammation in Mice

    doi: 10.1371/journal.pone.0126599

    Figure Lengend Snippet: Quantitative PCR of microarray genes. Selected qPCR of genes that were 1.5 fold or more changed by RSV treatment in the microarray (RSV treated compared with the IMQ group). The mRNA levels were quantified using MYO18B as reference gene. (a) Phosphoenolpyruvate Carboxykinase 1 (PCK1). (b) Tripartite Motif Containing 63, E3 Ubiquitin Protein Ligase (TRIM63). (c) Protein Phosphatase 1, Regulatory (inhibitor) Subunit 3C (PPP1R3C). Columns in (a-c) are group means ±SEM (n = 8, n = 10, n = 10 for controls, IMQ, IMQ-RSV respectively). Clamped bar with * above indicates the pair of column means are significantly different (p

    Article Snippet: The PCR reactions were performed in duplicates using the KAPA SYBR FAST qPCR kit (Kapa Biosystems, Inc., Woburn, MA) in a LightCycler 480 (Roche Applied Science) using the following protocol: One step at 95°C for 3 min, then 95°C for 10 s, 60°C for 20 s, and 72°C for 10 s. The increase in fluorescence was measured in real time during the extension step and a final melt curve analysis was performed to verify the specificity of the amplification.

    Techniques: Real-time Polymerase Chain Reaction, Microarray

    ME1 knockdown mimicked the inhibitory effect of miR-612 overexpression in bladder cancer cells. (A) mRNA and (B) protein levels of ME1 in T24 cells following transfection with si-ME1 or si-NC were measured using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. GAPDH and β-actin were used as internal controls, respectively. (C) Cell proliferation, (D) colony formation, (E) Transwell tumor cell migration assay and (F) Transwell tumor cell invasion assays in bladder cancer T24 cells transfected with si-ME1 or si-NC. (G) Protein levels of EMT-associated markers in T24 cells transfected with si-ME1 or si-NC were evaluated by western blot analysis. * P

    Journal: International Journal of Oncology

    Article Title: Tumor-suppressing effects of microRNA-612 in bladder cancer cells by targeting malic enzyme 1 expression

    doi: 10.3892/ijo.2018.4342

    Figure Lengend Snippet: ME1 knockdown mimicked the inhibitory effect of miR-612 overexpression in bladder cancer cells. (A) mRNA and (B) protein levels of ME1 in T24 cells following transfection with si-ME1 or si-NC were measured using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. GAPDH and β-actin were used as internal controls, respectively. (C) Cell proliferation, (D) colony formation, (E) Transwell tumor cell migration assay and (F) Transwell tumor cell invasion assays in bladder cancer T24 cells transfected with si-ME1 or si-NC. (G) Protein levels of EMT-associated markers in T24 cells transfected with si-ME1 or si-NC were evaluated by western blot analysis. * P

    Article Snippet: The amplification procedure was as follows: 5 min at 95°C, followed by 40 cycles at 95°C for 30 sec and 65°C for 45 sec. For detection of the ME1 mRNA level, cDNA was synthesized using the PrimeScript RT Reagent kit (Takara Bio, Inc., Tokyo, Japan), according to the manufacturer's protocol. qPCR for mRNA detection was also performed using the KAPA SYBR FAST qPCR kit (Kapa Biosystems, Inc.), The primers used were as follows: ME1 forward, 5′-GCAGGTCTCCTTGCAGCTCT-3′ and reverse, 5′-TCCAAGGCCATCACA ATCAG-3′; GAPDH forward, 5′-ATGTCGTGGAGTCTACTGGC-3′ and reverse, 5′-TGACCTTGCCCACAGCCTTG-3′.

    Techniques: Over Expression, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Cell Migration Assay

    ME1 overexpression partially rescues the inhibitory effects of miR-612 expression on bladder cancer cells. (A) mRNA and (B) protein levels of ME1 in T24 cells co-transfected with miR-612 mimic or miR-NC, and with or without ME1 cDNA were measured using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. (C) Cell proliferation, (D) colony formation, (E) Transwell tumor cell migration and (F) Transwell tumor cell invasion assays in T24 cells co-transfected with miR-612 mimic or miR-NC, and with or without ME1 cDNA. * P

    Journal: International Journal of Oncology

    Article Title: Tumor-suppressing effects of microRNA-612 in bladder cancer cells by targeting malic enzyme 1 expression

    doi: 10.3892/ijo.2018.4342

    Figure Lengend Snippet: ME1 overexpression partially rescues the inhibitory effects of miR-612 expression on bladder cancer cells. (A) mRNA and (B) protein levels of ME1 in T24 cells co-transfected with miR-612 mimic or miR-NC, and with or without ME1 cDNA were measured using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. (C) Cell proliferation, (D) colony formation, (E) Transwell tumor cell migration and (F) Transwell tumor cell invasion assays in T24 cells co-transfected with miR-612 mimic or miR-NC, and with or without ME1 cDNA. * P

    Article Snippet: The amplification procedure was as follows: 5 min at 95°C, followed by 40 cycles at 95°C for 30 sec and 65°C for 45 sec. For detection of the ME1 mRNA level, cDNA was synthesized using the PrimeScript RT Reagent kit (Takara Bio, Inc., Tokyo, Japan), according to the manufacturer's protocol. qPCR for mRNA detection was also performed using the KAPA SYBR FAST qPCR kit (Kapa Biosystems, Inc.), The primers used were as follows: ME1 forward, 5′-GCAGGTCTCCTTGCAGCTCT-3′ and reverse, 5′-TCCAAGGCCATCACA ATCAG-3′; GAPDH forward, 5′-ATGTCGTGGAGTCTACTGGC-3′ and reverse, 5′-TGACCTTGCCCACAGCCTTG-3′.

    Techniques: Over Expression, Expressing, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Migration

    ME1 is a direct target of miR-612. (A) Bioinformatic analysis indicated that ME1 is the target of miR-612. The presumable miR-612-binding sites in the 3′-UTR of ME1 cDNA and the mutant binding sites are shown. (B) The relative luciferase activity was determined in T24 cells following co-transfection with a luciferase reporter plasmid (WT or MUT 3′-UTR ME1 cDNA), and miR-612 mimic or miR-NC. (C) mRNA and (D) protein levels of ME1 in T24 cells following transfection with miR-612 mimic or miR-NC were analyzed using RT-qPCR and western blot analysis, respectively. GAPDH and β-actin served as the internal controls, respectively. (E) The levels of ME1 mRNA were measured using RT-qPCR in 46 paired bladder cancer and ANT specimens. GAPDH was used as an internal control. (F) The correlation between ME1 and miR-612 expression in bladder cancer tissue samples (n=46) was analyzed using Spearman's correlation test. ** P

    Journal: International Journal of Oncology

    Article Title: Tumor-suppressing effects of microRNA-612 in bladder cancer cells by targeting malic enzyme 1 expression

    doi: 10.3892/ijo.2018.4342

    Figure Lengend Snippet: ME1 is a direct target of miR-612. (A) Bioinformatic analysis indicated that ME1 is the target of miR-612. The presumable miR-612-binding sites in the 3′-UTR of ME1 cDNA and the mutant binding sites are shown. (B) The relative luciferase activity was determined in T24 cells following co-transfection with a luciferase reporter plasmid (WT or MUT 3′-UTR ME1 cDNA), and miR-612 mimic or miR-NC. (C) mRNA and (D) protein levels of ME1 in T24 cells following transfection with miR-612 mimic or miR-NC were analyzed using RT-qPCR and western blot analysis, respectively. GAPDH and β-actin served as the internal controls, respectively. (E) The levels of ME1 mRNA were measured using RT-qPCR in 46 paired bladder cancer and ANT specimens. GAPDH was used as an internal control. (F) The correlation between ME1 and miR-612 expression in bladder cancer tissue samples (n=46) was analyzed using Spearman's correlation test. ** P

    Article Snippet: The amplification procedure was as follows: 5 min at 95°C, followed by 40 cycles at 95°C for 30 sec and 65°C for 45 sec. For detection of the ME1 mRNA level, cDNA was synthesized using the PrimeScript RT Reagent kit (Takara Bio, Inc., Tokyo, Japan), according to the manufacturer's protocol. qPCR for mRNA detection was also performed using the KAPA SYBR FAST qPCR kit (Kapa Biosystems, Inc.), The primers used were as follows: ME1 forward, 5′-GCAGGTCTCCTTGCAGCTCT-3′ and reverse, 5′-TCCAAGGCCATCACA ATCAG-3′; GAPDH forward, 5′-ATGTCGTGGAGTCTACTGGC-3′ and reverse, 5′-TGACCTTGCCCACAGCCTTG-3′.

    Techniques: Binding Assay, Mutagenesis, Luciferase, Activity Assay, Cotransfection, Plasmid Preparation, Transfection, Quantitative RT-PCR, Western Blot, Expressing

    Real-time PCR and Western blotting analyses of CLIC2, CLIC3 and CLIC4 in ESCCs and paired NTs. (A-C): CLIC2 (A), CLIC3 (B) and CLIC4 (C) mRNA assays by qPCR between ESCCs and paired NTs. (D-F): Representative CLIC2 (D), CLIC3 (E) and CLIC4 (F) protein assays by Western blotting in ESCCs and paired NTs, GAPDH served as loading control. ( G-I): Statistical analyses of CLIC2 (G), CLIC3 (H) and CLIC4 (I) protein levels between ESCCs and paired NTs.

    Journal: Journal of Cancer

    Article Title: Analysis of Differentially Expressed Genes in a Chinese Cohort of Esophageal Squamous Cell Carcinoma

    doi: 10.7150/jca.40850

    Figure Lengend Snippet: Real-time PCR and Western blotting analyses of CLIC2, CLIC3 and CLIC4 in ESCCs and paired NTs. (A-C): CLIC2 (A), CLIC3 (B) and CLIC4 (C) mRNA assays by qPCR between ESCCs and paired NTs. (D-F): Representative CLIC2 (D), CLIC3 (E) and CLIC4 (F) protein assays by Western blotting in ESCCs and paired NTs, GAPDH served as loading control. ( G-I): Statistical analyses of CLIC2 (G), CLIC3 (H) and CLIC4 (I) protein levels between ESCCs and paired NTs.

    Article Snippet: For real-time PCR, 1 μl of the cDNA was used for mRNA amplification using KAPA SYBR FAST PCR Universal Kit (KapaBio systems) in a Mini-Opticon Thermal Cycler (BIORAD).

    Techniques: Real-time Polymerase Chain Reaction, Western Blot