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    Name:
    Small RNA Cloning Kit
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    Catalog Number:
    rr065
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    Size:
    10 Rxns
    Category:
    cDNA synthesis kits cDNA synthesis
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    Structured Review

    TaKaRa cdna synthesis kit
    Variation in expression level of ATP5b in muscle and heart tissues of exercised and BCAAs supplemented mice. <t>qRT-PCR</t> on <t>cDNA</t> samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of ATP5b transcript level in 20BCAA group ( p

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    Images

    1) Product Images from "PPARγ/Pgc-1α-Fndc5 pathway up-regulation in gastrocnemius and heart muscle of exercised, branched chain amino acid diet fed mice"

    Article Title: PPARγ/Pgc-1α-Fndc5 pathway up-regulation in gastrocnemius and heart muscle of exercised, branched chain amino acid diet fed mice

    Journal: Nutrition & Metabolism

    doi: 10.1186/s12986-018-0298-3

    Variation in expression level of ATP5b in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of ATP5b transcript level in 20BCAA group ( p
    Figure Legend Snippet: Variation in expression level of ATP5b in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of ATP5b transcript level in 20BCAA group ( p

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Fndc5 transcript levels were increased in heart and muscle tissues of exercised and BCAAs supplemented mice. As elucidated, qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase in Fndc5 transcript level ( p
    Figure Legend Snippet: Fndc5 transcript levels were increased in heart and muscle tissues of exercised and BCAAs supplemented mice. As elucidated, qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase in Fndc5 transcript level ( p

    Techniques Used: Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Modulation in relative expression levels of ATP5a1 in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of ATP5a1 transcript level in 20BCAA group ( p
    Figure Legend Snippet: Modulation in relative expression levels of ATP5a1 in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of ATP5a1 transcript level in 20BCAA group ( p

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Variation in relative expression levels of Cox4i1 in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of Cox4i1 transcript level in in 20BCAA group ( p
    Figure Legend Snippet: Variation in relative expression levels of Cox4i1 in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of Cox4i1 transcript level in in 20BCAA group ( p

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Modulation in transcript level of PPARγ in gastrocnemius muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and 20 mg/mL BCAAs supplemented mice (20BCAA/Ex) indicated a significant increase in PPARγ transcript level ( p
    Figure Legend Snippet: Modulation in transcript level of PPARγ in gastrocnemius muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and 20 mg/mL BCAAs supplemented mice (20BCAA/Ex) indicated a significant increase in PPARγ transcript level ( p

    Techniques Used: Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Expression levels of Pgc-1α was increased in heart and muscle tissues of exercised and 20 mg/mL BCAAs (20BCAA/Ex) supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAA supplemented mice indicated most increase of Pgc-1α transcript level in 20BCAA group (p
    Figure Legend Snippet: Expression levels of Pgc-1α was increased in heart and muscle tissues of exercised and 20 mg/mL BCAAs (20BCAA/Ex) supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAA supplemented mice indicated most increase of Pgc-1α transcript level in 20BCAA group (p

    Techniques Used: Expressing, Pyrolysis Gas Chromatography, Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Modulation in transcript levels of Tfam in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a most increase of Tfam transcript level in 20BCAA group ( p
    Figure Legend Snippet: Modulation in transcript levels of Tfam in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a most increase of Tfam transcript level in 20BCAA group ( p

    Techniques Used: Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Modulation in relative expression level of Sirt1 in gastrocnemius and heart muscles of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and 20 mg/mL BCAAs supplemented mice (20BCAA/Ex) indicated a significant increase of Sirt1 transcript level ( p
    Figure Legend Snippet: Modulation in relative expression level of Sirt1 in gastrocnemius and heart muscles of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and 20 mg/mL BCAAs supplemented mice (20BCAA/Ex) indicated a significant increase of Sirt1 transcript level ( p

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR, Derivative Assay

    2) Product Images from "Different Involvement of Promoter Methylation in the Expression of Organic Cation/Carnitine Transporter 2 (OCTN2) in Cancer Cell Lines"

    Article Title: Different Involvement of Promoter Methylation in the Expression of Organic Cation/Carnitine Transporter 2 (OCTN2) in Cancer Cell Lines

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0076474

    Effects of DCA on mRNA and protein levels of OCTN2 in cancer cell lines. (A), analysis of mRNA levels of OCTN2 in DCA-treated cancer cell lines. After treatment with 0.5 and 1 μM DCA, total RNA in cancer cells was purified by TRIzol reagent for cDNA synthesis and the mRNA levels of OCTN2 were analyzed by Quantitative RT-PCR. (B), analysis of protein levels of OCTN2 in DCA-treated cancer cell lines. Total proteins were extracted and protein levels of OCTN2 were analyzed by western blotting. The relative expression of mRNA and protein was normalized to β-actin. Data presented represent the mean ± S.D. of three independent experiments. Significant difference from control group of 0.1% DMSO is denoted with asterisks (*, p
    Figure Legend Snippet: Effects of DCA on mRNA and protein levels of OCTN2 in cancer cell lines. (A), analysis of mRNA levels of OCTN2 in DCA-treated cancer cell lines. After treatment with 0.5 and 1 μM DCA, total RNA in cancer cells was purified by TRIzol reagent for cDNA synthesis and the mRNA levels of OCTN2 were analyzed by Quantitative RT-PCR. (B), analysis of protein levels of OCTN2 in DCA-treated cancer cell lines. Total proteins were extracted and protein levels of OCTN2 were analyzed by western blotting. The relative expression of mRNA and protein was normalized to β-actin. Data presented represent the mean ± S.D. of three independent experiments. Significant difference from control group of 0.1% DMSO is denoted with asterisks (*, p

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

    Expression of OCTN2 in cancer cell lines. (A), analysis of mRNA levels of OCTN2 in cancer cell lines. Total RNA in cancer cells was purified by TRIzol reagent for cDNA synthesis and the mRNA levels of OCTN2 were analyzed by Quantitative RT-PCR. (B), analysis of protein levels of OCTN2 in cancer cells. Total proteins were extracted and protein levels of OCTN2 were analyzed by western blotting. The relative expression of mRNA and protein was normalized to β-actin. The expression of HepG2 was set as 1. All of mRNA and protein analysis were performed three times. Significant difference from HepG2 or LS174T cell is denoted with asterisk (*, p
    Figure Legend Snippet: Expression of OCTN2 in cancer cell lines. (A), analysis of mRNA levels of OCTN2 in cancer cell lines. Total RNA in cancer cells was purified by TRIzol reagent for cDNA synthesis and the mRNA levels of OCTN2 were analyzed by Quantitative RT-PCR. (B), analysis of protein levels of OCTN2 in cancer cells. Total proteins were extracted and protein levels of OCTN2 were analyzed by western blotting. The relative expression of mRNA and protein was normalized to β-actin. The expression of HepG2 was set as 1. All of mRNA and protein analysis were performed three times. Significant difference from HepG2 or LS174T cell is denoted with asterisk (*, p

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

    3) Product Images from "Sulforaphene Suppresses Adipocyte Differentiation via Induction of Post-Translational Degradation of CCAAT/Enhancer Binding Protein Beta (C/EBPβ)"

    Article Title: Sulforaphene Suppresses Adipocyte Differentiation via Induction of Post-Translational Degradation of CCAAT/Enhancer Binding Protein Beta (C/EBPβ)

    Journal: Nutrients

    doi: 10.3390/nu12030758

    Effect of sulforaphene (SFEN) on differentiation medium-induced C/EBPβ expression in 3T3-L1 pre-adipocytes. ( A ) Western blot analysis indicated that SFEN attenuated the protein expression levels of C/EBPβ (35 kDa) and β-actin (43 kDa) as housekeeping gene in a dose-dependent and ( B ) time-dependent manner. ( C ) The gene expression levels of C/ebpβ and β-actin as housekeeping gene in 3T3-L1 pre-adipocytes were determined using real-time PCR. 3T3-L1 pre-adipocytes were seeded in 6 cm dishes at a density of 0.75 × 10 4 cells per cm 2 . Confluent cells were incubated for two days in mouse adipocyte differentiation medium (MDM) with or without SFEN at a concentration of 5 or 10 μΜ. ( A , C ) At 48 h and ( B ) 4, 12, 24, and 48 h after the addition of MDM, ( A , B ) the cells were harvested by using 80 μL of RIPA buffer per dish to prepare samples for western blotting to determine the expression level of C/EBPβ. 40 μg of protein was loaded per lane on 10% gel. ( C ) The mature adipocytes were harvested for qPCR and 1 μg of RNA was used for the synthesis of cDNA. Data were obtained from three independent experiments. For Figure 4 B, similar data were obtained from an independent repeated experiment. Data are expressed as the means ± standard deviation ( n = 3). ##, significant difference between the control and the MDM control by ( p
    Figure Legend Snippet: Effect of sulforaphene (SFEN) on differentiation medium-induced C/EBPβ expression in 3T3-L1 pre-adipocytes. ( A ) Western blot analysis indicated that SFEN attenuated the protein expression levels of C/EBPβ (35 kDa) and β-actin (43 kDa) as housekeeping gene in a dose-dependent and ( B ) time-dependent manner. ( C ) The gene expression levels of C/ebpβ and β-actin as housekeeping gene in 3T3-L1 pre-adipocytes were determined using real-time PCR. 3T3-L1 pre-adipocytes were seeded in 6 cm dishes at a density of 0.75 × 10 4 cells per cm 2 . Confluent cells were incubated for two days in mouse adipocyte differentiation medium (MDM) with or without SFEN at a concentration of 5 or 10 μΜ. ( A , C ) At 48 h and ( B ) 4, 12, 24, and 48 h after the addition of MDM, ( A , B ) the cells were harvested by using 80 μL of RIPA buffer per dish to prepare samples for western blotting to determine the expression level of C/EBPβ. 40 μg of protein was loaded per lane on 10% gel. ( C ) The mature adipocytes were harvested for qPCR and 1 μg of RNA was used for the synthesis of cDNA. Data were obtained from three independent experiments. For Figure 4 B, similar data were obtained from an independent repeated experiment. Data are expressed as the means ± standard deviation ( n = 3). ##, significant difference between the control and the MDM control by ( p

    Techniques Used: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Incubation, Concentration Assay, Standard Deviation

    Effects of sulforaphene on differentiation medium-induced adipogenesis and the expression of two adipogenic genes—PPARγ and C/EBPα—in 3T3-L1 pre-adipocytes. ( A ) Image of differentiated 3T3-L1 adipocytes stained with Oil Red O (Magnification: 200×) ( B ) Quantification of intracellular lipid accumulation in differentiated 3T3-L1 adipocytes. ( C ) Western blot analysis of the protein expression levels of PPARγ (54, 57 kDa), C/EBPα (28, 42 kDa), and β-actin (43 kDa) as a housekeeping gene. Quantitative PCR analysis of the gene expression levels of ( D ) Pparγ , ( E ) C/ebpα , and β-actin as housekeeping gene. ( A , B ) 3T3-L1 pre-adipocytes were seeded at a density of 1.25 × 10 4 cells per cm 2 in 24-well plates and ( C – E ) 0.75 × 10 4 cells per cm 2 in 6 cm dishes. Confluent cells were incubated for two days in mouse adipocyte differentiation medium (MDM) with or without SFEN treatment at a concentration of 5 or 10 μΜ. After six days of differentiation, ( A ) the cell layer was stained with Oil Red O and ( B ) intracellular lipid accumulation was quantified. ( C ) The mature adipocytes were harvested by using 80 μL of RIPA buffer per dish for western blotting. 40 μg of protein was loaded per lane on 10% gel. ( D , E ) The mature adipocytes were harvested for qPCR and 1 μg of RNA was used for the synthesis of cDNA. Data were obtained from three independent experiments and are expressed as the means ± standard deviation ( n = 3). ##, significant difference ( p
    Figure Legend Snippet: Effects of sulforaphene on differentiation medium-induced adipogenesis and the expression of two adipogenic genes—PPARγ and C/EBPα—in 3T3-L1 pre-adipocytes. ( A ) Image of differentiated 3T3-L1 adipocytes stained with Oil Red O (Magnification: 200×) ( B ) Quantification of intracellular lipid accumulation in differentiated 3T3-L1 adipocytes. ( C ) Western blot analysis of the protein expression levels of PPARγ (54, 57 kDa), C/EBPα (28, 42 kDa), and β-actin (43 kDa) as a housekeeping gene. Quantitative PCR analysis of the gene expression levels of ( D ) Pparγ , ( E ) C/ebpα , and β-actin as housekeeping gene. ( A , B ) 3T3-L1 pre-adipocytes were seeded at a density of 1.25 × 10 4 cells per cm 2 in 24-well plates and ( C – E ) 0.75 × 10 4 cells per cm 2 in 6 cm dishes. Confluent cells were incubated for two days in mouse adipocyte differentiation medium (MDM) with or without SFEN treatment at a concentration of 5 or 10 μΜ. After six days of differentiation, ( A ) the cell layer was stained with Oil Red O and ( B ) intracellular lipid accumulation was quantified. ( C ) The mature adipocytes were harvested by using 80 μL of RIPA buffer per dish for western blotting. 40 μg of protein was loaded per lane on 10% gel. ( D , E ) The mature adipocytes were harvested for qPCR and 1 μg of RNA was used for the synthesis of cDNA. Data were obtained from three independent experiments and are expressed as the means ± standard deviation ( n = 3). ##, significant difference ( p

    Techniques Used: Expressing, Staining, Western Blot, Real-time Polymerase Chain Reaction, Incubation, Concentration Assay, Standard Deviation

    4) Product Images from "Low Toxicity of Deoxynivalenol-3-Glucoside in Microbial Cells"

    Article Title: Low Toxicity of Deoxynivalenol-3-Glucoside in Microbial Cells

    Journal: Toxins

    doi: 10.3390/toxins7010187

    Semi-quantitative analysis of DNA microarray data. Total RNA that was used in the DNA microarray analysis and DON-treated RNA samples were prepared for synthesizing cDNA templates. Bars = SE; n = 3.
    Figure Legend Snippet: Semi-quantitative analysis of DNA microarray data. Total RNA that was used in the DNA microarray analysis and DON-treated RNA samples were prepared for synthesizing cDNA templates. Bars = SE; n = 3.

    Techniques Used: Microarray

    5) Product Images from "Meiotic spindle stability depends on MAPK-interacting and spindle-stabilizing protein (MISS), a new MAPK substrate"

    Article Title: Meiotic spindle stability depends on MAPK-interacting and spindle-stabilizing protein (MISS), a new MAPK substrate

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.200202052

    Interaction between MISS and MAPK. (A) Two-hybrid interaction between MISS and MAPK. MISS interacts with ERK2WT and ERK2KD but not with a negative control. The yeast transformed with LexA-ERK2WT, LexA-ERK2KD ( Waskiewicz et al., 1997 ), LexA-53 (positive control; CLONTECH Laboratories, Inc.), LexA-Su(Fu), or LexA alone were mated with yeast transformed respectively with B42-cDNA, B42-AD-T (positive control; CLONTECH Laboratories, Inc.), or empty B42. The diploids obtained were assayed for transactivation of both the β-galactosidase and the LEU2 reporter genes on glucose (Glu)- or galactose (Gal/Raf)-containing mediums. The B42 fusion proteins are under the control of the galactose promoter. The B42-cDNA clearly interacts both with fusions of ERK2WT and ERK2KD at levels identical to the positive control, whereas it does not interact with a negative control (Su[Fu]). (B) MISS coimmunoprecipitates with endogenous p42 mpk from Xenopus oocyte extracts. Oocyte extracts expressing Myc–Wnt11 ( Xenopus Wint11; negative control) or Myc–MISS RNA (lanes 1 and 2). Anti-p42 mpk1 immunoprecipitates (lanes 3 and 4) prepared from the oocyte lysates were analyzed by immunoblotting with anti-Myc antibody. This experiment was performed three times. (C) Expression of MISS RNA in immature oocytes and two cell stage embryos. For RT-PCR analysis, total RNA was isolated from mouse ovaries (lane 2), immature oocytes (lanes 3 and 4), and two cell stage embryos (lanes 5 and 6) and treated with or without reverse transcriptase (RT, + or −). Lane 1 corresponds to a PCR control (water). (D) MISS accumulates during meiotic maturation. 2,000 immature (lane 1) or mature (lane 2) oocytes were collected and immunoblotted using an affinity-purified anti-MISS antibody.
    Figure Legend Snippet: Interaction between MISS and MAPK. (A) Two-hybrid interaction between MISS and MAPK. MISS interacts with ERK2WT and ERK2KD but not with a negative control. The yeast transformed with LexA-ERK2WT, LexA-ERK2KD ( Waskiewicz et al., 1997 ), LexA-53 (positive control; CLONTECH Laboratories, Inc.), LexA-Su(Fu), or LexA alone were mated with yeast transformed respectively with B42-cDNA, B42-AD-T (positive control; CLONTECH Laboratories, Inc.), or empty B42. The diploids obtained were assayed for transactivation of both the β-galactosidase and the LEU2 reporter genes on glucose (Glu)- or galactose (Gal/Raf)-containing mediums. The B42 fusion proteins are under the control of the galactose promoter. The B42-cDNA clearly interacts both with fusions of ERK2WT and ERK2KD at levels identical to the positive control, whereas it does not interact with a negative control (Su[Fu]). (B) MISS coimmunoprecipitates with endogenous p42 mpk from Xenopus oocyte extracts. Oocyte extracts expressing Myc–Wnt11 ( Xenopus Wint11; negative control) or Myc–MISS RNA (lanes 1 and 2). Anti-p42 mpk1 immunoprecipitates (lanes 3 and 4) prepared from the oocyte lysates were analyzed by immunoblotting with anti-Myc antibody. This experiment was performed three times. (C) Expression of MISS RNA in immature oocytes and two cell stage embryos. For RT-PCR analysis, total RNA was isolated from mouse ovaries (lane 2), immature oocytes (lanes 3 and 4), and two cell stage embryos (lanes 5 and 6) and treated with or without reverse transcriptase (RT, + or −). Lane 1 corresponds to a PCR control (water). (D) MISS accumulates during meiotic maturation. 2,000 immature (lane 1) or mature (lane 2) oocytes were collected and immunoblotted using an affinity-purified anti-MISS antibody.

    Techniques Used: Negative Control, Transformation Assay, Positive Control, Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation, Polymerase Chain Reaction, Affinity Purification

    6) Product Images from "Post-duplication charge evolution of phosphoglucose isomerases in teleost fishes through weak selection on many amino acid sites"

    Article Title: Post-duplication charge evolution of phosphoglucose isomerases in teleost fishes through weak selection on many amino acid sites

    Journal: BMC Evolutionary Biology

    doi: 10.1186/1471-2148-7-204

    Molecular phylogeny and spatial expression patterns of Pgi . (A) Bayesian tree of Pgi genes derived from 20 vertebrates. Numbers indicate percent posterior probabilities for the Bayesian tree (left) and bootstrap support values by the maximum likelihood method (right). Arrow denotes a gene duplication event. In cDNA clones, only one Pgi was identified from non-teleosts, whereas two Pgi genes were identified from teleosts. The two Pgi genes differed by about 20% in amino acid sequence, and were grouped into separate clades ( Pgi-1 and Pgi-2 ). In both clades, the gene relationships were consistent with the evolutionary relationships of teleost species [18, 19, 21]. (B) Partial-length gel images of the RT-PCR expression analysis of Pgi genes and positive control ( β-actin ) genes in ray-finned fishes. The tree in the left panel shows the relationships among the Pgi genes inferred in this study. The black circle on the tree denotes the timing of the Pgi gene duplication event. Letters indicate tissues: M, muscle; L, liver; H, heart; Gi, gill; B, brain; K, kidney. Full-length gels, including negative controls and size markers, are presented in Additional file 1 : Fig. S5.
    Figure Legend Snippet: Molecular phylogeny and spatial expression patterns of Pgi . (A) Bayesian tree of Pgi genes derived from 20 vertebrates. Numbers indicate percent posterior probabilities for the Bayesian tree (left) and bootstrap support values by the maximum likelihood method (right). Arrow denotes a gene duplication event. In cDNA clones, only one Pgi was identified from non-teleosts, whereas two Pgi genes were identified from teleosts. The two Pgi genes differed by about 20% in amino acid sequence, and were grouped into separate clades ( Pgi-1 and Pgi-2 ). In both clades, the gene relationships were consistent with the evolutionary relationships of teleost species [18, 19, 21]. (B) Partial-length gel images of the RT-PCR expression analysis of Pgi genes and positive control ( β-actin ) genes in ray-finned fishes. The tree in the left panel shows the relationships among the Pgi genes inferred in this study. The black circle on the tree denotes the timing of the Pgi gene duplication event. Letters indicate tissues: M, muscle; L, liver; H, heart; Gi, gill; B, brain; K, kidney. Full-length gels, including negative controls and size markers, are presented in Additional file 1 : Fig. S5.

    Techniques Used: Expressing, Derivative Assay, Clone Assay, Sequencing, Reverse Transcription Polymerase Chain Reaction, Positive Control

    7) Product Images from "PPARγ/Pgc-1α-Fndc5 pathway up-regulation in gastrocnemius and heart muscle of exercised, branched chain amino acid diet fed mice"

    Article Title: PPARγ/Pgc-1α-Fndc5 pathway up-regulation in gastrocnemius and heart muscle of exercised, branched chain amino acid diet fed mice

    Journal: Nutrition & Metabolism

    doi: 10.1186/s12986-018-0298-3

    Variation in expression level of ATP5b in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of ATP5b transcript level in 20BCAA group ( p
    Figure Legend Snippet: Variation in expression level of ATP5b in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of ATP5b transcript level in 20BCAA group ( p

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Modulation in relative expression levels of ATP5a1 in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of ATP5a1 transcript level in 20BCAA group ( p
    Figure Legend Snippet: Modulation in relative expression levels of ATP5a1 in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of ATP5a1 transcript level in 20BCAA group ( p

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Fndc5 transcript levels were increased in heart and muscle tissues of exercised and BCAAs supplemented mice. As elucidated, qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase in Fndc5 transcript level ( p
    Figure Legend Snippet: Fndc5 transcript levels were increased in heart and muscle tissues of exercised and BCAAs supplemented mice. As elucidated, qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase in Fndc5 transcript level ( p

    Techniques Used: Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Variation in relative expression levels of Cox4i1 in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of Cox4i1 transcript level in in 20BCAA group ( p
    Figure Legend Snippet: Variation in relative expression levels of Cox4i1 in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a significant increase of Cox4i1 transcript level in in 20BCAA group ( p

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Modulation in transcript level of PPARγ in gastrocnemius muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and 20 mg/mL BCAAs supplemented mice (20BCAA/Ex) indicated a significant increase in PPARγ transcript level ( p
    Figure Legend Snippet: Modulation in transcript level of PPARγ in gastrocnemius muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and 20 mg/mL BCAAs supplemented mice (20BCAA/Ex) indicated a significant increase in PPARγ transcript level ( p

    Techniques Used: Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Expression levels of Pgc-1α was increased in heart and muscle tissues of exercised and 20 mg/mL BCAAs (20BCAA/Ex) supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAA supplemented mice indicated most increase of Pgc-1α transcript level in 20BCAA group (p
    Figure Legend Snippet: Expression levels of Pgc-1α was increased in heart and muscle tissues of exercised and 20 mg/mL BCAAs (20BCAA/Ex) supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAA supplemented mice indicated most increase of Pgc-1α transcript level in 20BCAA group (p

    Techniques Used: Expressing, Pyrolysis Gas Chromatography, Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Modulation in transcript levels of Tfam in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a most increase of Tfam transcript level in 20BCAA group ( p
    Figure Legend Snippet: Modulation in transcript levels of Tfam in muscle and heart tissues of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and BCAAs supplemented mice indicated a most increase of Tfam transcript level in 20BCAA group ( p

    Techniques Used: Mouse Assay, Quantitative RT-PCR, Derivative Assay

    Modulation in relative expression level of Sirt1 in gastrocnemius and heart muscles of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and 20 mg/mL BCAAs supplemented mice (20BCAA/Ex) indicated a significant increase of Sirt1 transcript level ( p
    Figure Legend Snippet: Modulation in relative expression level of Sirt1 in gastrocnemius and heart muscles of exercised and BCAAs supplemented mice. qRT-PCR on cDNA samples of gastrocnemius muscle ( a ) and heart ( b ) tissues derived from exercised and 20 mg/mL BCAAs supplemented mice (20BCAA/Ex) indicated a significant increase of Sirt1 transcript level ( p

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR, Derivative Assay

    8) Product Images from "Role of gob-5 in mucus overproduction and airway hyperresponsiveness in asthma"

    Article Title: Role of gob-5 in mucus overproduction and airway hyperresponsiveness in asthma

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

    doi: 10.1073/pnas.081510898

    Selective expression of gob-5 mRNA in airway epithelia from AHR-model mice. ( A ) Tissue distribution of gob-5 was determined by Northern blot analysis in normal and AHR-model mice. ( Upper ) The results obtained by hybridization with a gob-5 cDNA probe. The murine tissues from which the poly(A) + RNA was prepared are indicated at the top. The same filter was rehybridized with a β -actin probe to control for equal loading ( Lower ). ( B , C , E , and F ) Lung sections of normal ( B and C ) and AHR-model mice ( E and F ) were in situ hybridized with DIG-labeled, single-stranded antisense ( B and E ) and sense ( C and F ) RNA probes. ( D and G ) The same lung sections of normal ( D ) and AHR-model mice ( G ) were stained with PAS and counterstained with hematoxylin. Original magnification, ×200. Three different experiments showed similar results.
    Figure Legend Snippet: Selective expression of gob-5 mRNA in airway epithelia from AHR-model mice. ( A ) Tissue distribution of gob-5 was determined by Northern blot analysis in normal and AHR-model mice. ( Upper ) The results obtained by hybridization with a gob-5 cDNA probe. The murine tissues from which the poly(A) + RNA was prepared are indicated at the top. The same filter was rehybridized with a β -actin probe to control for equal loading ( Lower ). ( B , C , E , and F ) Lung sections of normal ( B and C ) and AHR-model mice ( E and F ) were in situ hybridized with DIG-labeled, single-stranded antisense ( B and E ) and sense ( C and F ) RNA probes. ( D and G ) The same lung sections of normal ( D ) and AHR-model mice ( G ) were stained with PAS and counterstained with hematoxylin. Original magnification, ×200. Three different experiments showed similar results.

    Techniques Used: Expressing, Mouse Assay, Northern Blot, Hybridization, In Situ, Labeling, Staining

    Mucus production in human mucoepidermoid cells transfected with g ob-5 or hCLCA1. ( A ) PAS staining was performed on NCI-H292 cells after transfection with control vector (pcDNA3.1), gob-5 expression vector (pcDNA-gob-5), or hCLCA1 expression vector (pcDNA-hCLCA1). ( B ) Induction of MUC5AC gene expression by transfection with gob-5 or hCLCA1 genes in NCI-H292 cells. Total RNA was extracted from transfected cells 3 days after transfection, and reverse transcription (RT)-PCR amplification was performed for hCLCA1 , gob-5 , and MUC5AC gene detection. Amplification of the GAPDH gene was used as a control for RNA loading. cDNA reactions in which the RT was omitted (− lanes) were used as negative controls for each condition. These data are representative of three independent experiments.
    Figure Legend Snippet: Mucus production in human mucoepidermoid cells transfected with g ob-5 or hCLCA1. ( A ) PAS staining was performed on NCI-H292 cells after transfection with control vector (pcDNA3.1), gob-5 expression vector (pcDNA-gob-5), or hCLCA1 expression vector (pcDNA-hCLCA1). ( B ) Induction of MUC5AC gene expression by transfection with gob-5 or hCLCA1 genes in NCI-H292 cells. Total RNA was extracted from transfected cells 3 days after transfection, and reverse transcription (RT)-PCR amplification was performed for hCLCA1 , gob-5 , and MUC5AC gene detection. Amplification of the GAPDH gene was used as a control for RNA loading. cDNA reactions in which the RT was omitted (− lanes) were used as negative controls for each condition. These data are representative of three independent experiments.

    Techniques Used: Transfection, Staining, Plasmid Preparation, Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification

    9) Product Images from "Circular RNA expression profiling reveals that circ-PLXNA1 functions in duck adipocyte differentiation"

    Article Title: Circular RNA expression profiling reveals that circ-PLXNA1 functions in duck adipocyte differentiation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0236069

    Characterization of circ-PLXNA1 in duck adipocyte. (A) The genomic loci of circ-PLXNA1 in PLXNA1 gene. (B) The validation strategy for circ-PLXNA1. Clear single bands were amplified from the cDNA of duck pre-adipocyte using divergent primers. Clear single bands could not be amplified from gDNA. (C) Sanger-Seq validated the head-to-tail junction region of duck pre-adipocyte circ-PLXNA1. (D) qRT–PCR result for the abundance of circ-PLXNA1 and PLXNA1 mRNA in duck adipocyte treated with RNase R. (E) The subcellular distribution of circ-PLXNA1. Nuclear and cytoplasmic RNA was extracted, and junction primers were used for circ-PLXNA1 detection. (F) The transcripts level of circ-PLXNA1 and PLXNA1 during duck adipocyte differentiation. The different letters show significant differences in common gene, P
    Figure Legend Snippet: Characterization of circ-PLXNA1 in duck adipocyte. (A) The genomic loci of circ-PLXNA1 in PLXNA1 gene. (B) The validation strategy for circ-PLXNA1. Clear single bands were amplified from the cDNA of duck pre-adipocyte using divergent primers. Clear single bands could not be amplified from gDNA. (C) Sanger-Seq validated the head-to-tail junction region of duck pre-adipocyte circ-PLXNA1. (D) qRT–PCR result for the abundance of circ-PLXNA1 and PLXNA1 mRNA in duck adipocyte treated with RNase R. (E) The subcellular distribution of circ-PLXNA1. Nuclear and cytoplasmic RNA was extracted, and junction primers were used for circ-PLXNA1 detection. (F) The transcripts level of circ-PLXNA1 and PLXNA1 during duck adipocyte differentiation. The different letters show significant differences in common gene, P

    Techniques Used: Amplification, Quantitative RT-PCR

    Related Articles

    Clone Assay:

    Article Title: Characterization of a mycovirus associated with the brown discoloration of edible mushroom, Flammulina velutipes
    Article Snippet: .. dsRNA sequence dsRNA extracted from the purified virions was cloned using the Takara small RNA Cloning Kit (Takara Bio) according to the manufacturer's instructions, with a slight modification: RevaTraAce (Toyobo, Osaka, Japan) was used for cDNA synthesis instead of the M-MLV RTase included in the kit, with the aim of generating a longer cDNA. .. The cDNA was cloned into the pGEM-T easy vector (Promega, Madison, WI, USA) according to the manufacturer's instructions and transformed into E. coli DH5α (Toyobo).

    RNA Extraction:

    Article Title: The Rice BZ1 Locus Is Required for Glycosylation of Arabinogalactan Proteins and Galactolipid and Plays a Role in both Mechanical Strength and Leaf Color
    Article Snippet: .. qRT-PCR MeasurementTotal RNA extraction was conducted using the Total RNA Extraction Kit (TaKaRa) and first-strand cDNA was synthesized with the PrimeScript RT Master Mix (TaKaRa). .. Real-time RT-PCR was performed as described previously (Li et al., ) by using SYBR Premix Ex TaqII (TaKaRa) on an Applied Biosystems QuanStudio 3 Real-Time PCR System.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Comparative Effect of Recombinant Shiga Toxin in Induction of Pro- and Anti-Apoptotic Markers and Inflammatory Cytokines in Epithelial and Monocytic Cells
    Article Snippet: .. Reverse Transcription-Polymerase Chain Reaction analysis For reverse transcription-polymerase chain reaction analysis (RT-PCR), total RNA was extracted using an RNA isolation kit (Takara Biotech Co. Japan) in accordance with the supplier’s protocols. .. The RT-PCR for each sample was performed in 50 μL of reaction mixture containing 1-10 μg of cellular RNA (O157, rStx and control cell) in a 50 μL reaction mixture, containing 2 µM of a forward primer, 2 μM of a reverse primer, 1 μM of each deoxynucleoside triphosphate, 30 μL double distilled water (DDW), 0.5 unit enzyme and 10 μL of buffer.

    Article Title: Identification and characterization of a novel pic gene cluster responsible for picolinic acid degradation in Alcaligenes faecalis JQ135
    Article Snippet: .. Total RNA was isolated using an RNA isolation kit (TaKaRa) and reverse transcription-PCR (RT-PCR) was carried out with a PrimeScript RT reagent kit (TaKaRa). ..

    Agarose Gel Electrophoresis:

    Article Title: Identification of microRNAs in the Toxigenic Dinoflagellate Alexandrium catenella by High-Throughput Illumina Sequencing and Bioinformatic Analysis
    Article Snippet: .. The small RNAs ( < 200 nt) were extracted from the samples following the manufacturer's instructions for the RNAiso for Small RNA kit(TaKaRa).The integrity and quality of sRNAs was evaluated by 3% agarose gel electrophoresis and the NanoDrop 2000 (Thermo, USA).Equal amounts of high-quality sRNAs (OD 260/280 ranging from 1.9 to 2.1) were reverse transcribed to cDNA. .. Each reverse transcriptase assay consisted of 10 μL of 2× miRNA Reaction Buffer Mix, 2μL of 0.1%BSA, 2μLof miRNA PrimeScript® RT Enzyme Mix, approximately 1μg of total RNA, and 5μL of RNase-free ddH2O.The resulting cDNA products were stored at −20°C.

    Synthesized:

    Article Title: The Rice BZ1 Locus Is Required for Glycosylation of Arabinogalactan Proteins and Galactolipid and Plays a Role in both Mechanical Strength and Leaf Color
    Article Snippet: .. qRT-PCR MeasurementTotal RNA extraction was conducted using the Total RNA Extraction Kit (TaKaRa) and first-strand cDNA was synthesized with the PrimeScript RT Master Mix (TaKaRa). .. Real-time RT-PCR was performed as described previously (Li et al., ) by using SYBR Premix Ex TaqII (TaKaRa) on an Applied Biosystems QuanStudio 3 Real-Time PCR System.

    Isolation:

    Article Title: Comparative Effect of Recombinant Shiga Toxin in Induction of Pro- and Anti-Apoptotic Markers and Inflammatory Cytokines in Epithelial and Monocytic Cells
    Article Snippet: .. Reverse Transcription-Polymerase Chain Reaction analysis For reverse transcription-polymerase chain reaction analysis (RT-PCR), total RNA was extracted using an RNA isolation kit (Takara Biotech Co. Japan) in accordance with the supplier’s protocols. .. The RT-PCR for each sample was performed in 50 μL of reaction mixture containing 1-10 μg of cellular RNA (O157, rStx and control cell) in a 50 μL reaction mixture, containing 2 µM of a forward primer, 2 μM of a reverse primer, 1 μM of each deoxynucleoside triphosphate, 30 μL double distilled water (DDW), 0.5 unit enzyme and 10 μL of buffer.

    Article Title: Identification and characterization of a novel pic gene cluster responsible for picolinic acid degradation in Alcaligenes faecalis JQ135
    Article Snippet: .. Total RNA was isolated using an RNA isolation kit (TaKaRa) and reverse transcription-PCR (RT-PCR) was carried out with a PrimeScript RT reagent kit (TaKaRa). ..

    Quantitative RT-PCR:

    Article Title: Mycobacterium bovis BCG Triggered MyD88 Induces miR-124 Feedback Negatively Regulates Immune Response in Alveolar Epithelial Cells
    Article Snippet: .. Quantitative reverse transcription-PCR (qRT-PCR) Small RNA was extracted from A549 cells or lung tissues using the RNAiso for small RNA kit, and total RNA was extracted using the AxyPrep Multisource Total RNA Miniprep kit per manufacturer's recommendations (Takara, Dalian, China). .. The RNA concentrations were determined using a NanoDrop instrument.

    Article Title: The Rice BZ1 Locus Is Required for Glycosylation of Arabinogalactan Proteins and Galactolipid and Plays a Role in both Mechanical Strength and Leaf Color
    Article Snippet: .. qRT-PCR MeasurementTotal RNA extraction was conducted using the Total RNA Extraction Kit (TaKaRa) and first-strand cDNA was synthesized with the PrimeScript RT Master Mix (TaKaRa). .. Real-time RT-PCR was performed as described previously (Li et al., ) by using SYBR Premix Ex TaqII (TaKaRa) on an Applied Biosystems QuanStudio 3 Real-Time PCR System.

    Purification:

    Article Title: Characterization of a mycovirus associated with the brown discoloration of edible mushroom, Flammulina velutipes
    Article Snippet: .. dsRNA sequence dsRNA extracted from the purified virions was cloned using the Takara small RNA Cloning Kit (Takara Bio) according to the manufacturer's instructions, with a slight modification: RevaTraAce (Toyobo, Osaka, Japan) was used for cDNA synthesis instead of the M-MLV RTase included in the kit, with the aim of generating a longer cDNA. .. The cDNA was cloned into the pGEM-T easy vector (Promega, Madison, WI, USA) according to the manufacturer's instructions and transformed into E. coli DH5α (Toyobo).

    Sequencing:

    Article Title: Characterization of a mycovirus associated with the brown discoloration of edible mushroom, Flammulina velutipes
    Article Snippet: .. dsRNA sequence dsRNA extracted from the purified virions was cloned using the Takara small RNA Cloning Kit (Takara Bio) according to the manufacturer's instructions, with a slight modification: RevaTraAce (Toyobo, Osaka, Japan) was used for cDNA synthesis instead of the M-MLV RTase included in the kit, with the aim of generating a longer cDNA. .. The cDNA was cloned into the pGEM-T easy vector (Promega, Madison, WI, USA) according to the manufacturer's instructions and transformed into E. coli DH5α (Toyobo).

    Modification:

    Article Title: Characterization of a mycovirus associated with the brown discoloration of edible mushroom, Flammulina velutipes
    Article Snippet: .. dsRNA sequence dsRNA extracted from the purified virions was cloned using the Takara small RNA Cloning Kit (Takara Bio) according to the manufacturer's instructions, with a slight modification: RevaTraAce (Toyobo, Osaka, Japan) was used for cDNA synthesis instead of the M-MLV RTase included in the kit, with the aim of generating a longer cDNA. .. The cDNA was cloned into the pGEM-T easy vector (Promega, Madison, WI, USA) according to the manufacturer's instructions and transformed into E. coli DH5α (Toyobo).

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    TaKaRa one step sybr primescript rt pcr kit ii
    Expression profiles of ALDH3 family members in human tissues. cDNAs from human liver, kidney, and brain tissue (human MTC (multiple tissue cDNA) panels, Takara Bio) ( A ) or prepared from primary keratinocytes (undifferentiated or differentiated for 7 days; CELLnTEC) ( B ) were subjected to <t>SYBR</t> Green-based real-time quantitative <t>PCR</t> using specific primers for ALDH3A1 , ALDH3A2 , ALDH3B1 , and GAPDH . Values represent the means ± S.D. from three independent reactions. A statistically significant difference is indicated (**, p
    One Step Sybr Primescript Rt Pcr Kit Ii, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 252 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Expression profiles of ALDH3 family members in human tissues. cDNAs from human liver, kidney, and brain tissue (human MTC (multiple tissue cDNA) panels, Takara Bio) ( A ) or prepared from primary keratinocytes (undifferentiated or differentiated for 7 days; CELLnTEC) ( B ) were subjected to SYBR Green-based real-time quantitative PCR using specific primers for ALDH3A1 , ALDH3A2 , ALDH3B1 , and GAPDH . Values represent the means ± S.D. from three independent reactions. A statistically significant difference is indicated (**, p

    Journal: The Journal of Biological Chemistry

    Article Title: Disruption of the Sjögren-Larsson Syndrome Gene Aldh3a2 in Mice Increases Keratinocyte Growth and Retards Skin Barrier Recovery *

    doi: 10.1074/jbc.M116.714030

    Figure Lengend Snippet: Expression profiles of ALDH3 family members in human tissues. cDNAs from human liver, kidney, and brain tissue (human MTC (multiple tissue cDNA) panels, Takara Bio) ( A ) or prepared from primary keratinocytes (undifferentiated or differentiated for 7 days; CELLnTEC) ( B ) were subjected to SYBR Green-based real-time quantitative PCR using specific primers for ALDH3A1 , ALDH3A2 , ALDH3B1 , and GAPDH . Values represent the means ± S.D. from three independent reactions. A statistically significant difference is indicated (**, p

    Article Snippet: Real-time quantitative PCR was performed using the One-Step SYBR PrimeScript RT-PCR Kit II (Takara Bio) on a CFX96 Touch real-time PCR detection system (Bio-Rad) according to the manufacturer's manual.

    Techniques: Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction

    Enhanced proliferation and oxidative stress response in Aldh3a2 −/− keratinocytes. A , primary keratinocytes were prepared from wild-type and Aldh3a2 KO mice and grown. Cell numbers were counted by microscopic observation. For each dish, cell numbers in three randomly chosen viewing fields were counted and summed. Values represent the means ± S.D. from four independent experiments. B , keratinocytes prepared from wild-type and Aldh3a2 KO mice were subjected to a [ 3 H]thymidine uptake assay. Values represent the means ± S.D. from three independent experiments. C and D , total RNAs prepared from wild-type and Aldh3a2 KO keratinocytes were subjected to SYBR Green-based real-time quantitative RT-PCR using specific primers for Ki67 ( C ), Hmox1 , Sod1 , Gclc , Gclm , Gsta1 , and Gapdh ( D ). Values are the amount of each mRNA relative to that of Gapdh and represent the means ± S.D. for three independent experiments. Statistically significant differences are indicated (**, p

    Journal: The Journal of Biological Chemistry

    Article Title: Disruption of the Sjögren-Larsson Syndrome Gene Aldh3a2 in Mice Increases Keratinocyte Growth and Retards Skin Barrier Recovery *

    doi: 10.1074/jbc.M116.714030

    Figure Lengend Snippet: Enhanced proliferation and oxidative stress response in Aldh3a2 −/− keratinocytes. A , primary keratinocytes were prepared from wild-type and Aldh3a2 KO mice and grown. Cell numbers were counted by microscopic observation. For each dish, cell numbers in three randomly chosen viewing fields were counted and summed. Values represent the means ± S.D. from four independent experiments. B , keratinocytes prepared from wild-type and Aldh3a2 KO mice were subjected to a [ 3 H]thymidine uptake assay. Values represent the means ± S.D. from three independent experiments. C and D , total RNAs prepared from wild-type and Aldh3a2 KO keratinocytes were subjected to SYBR Green-based real-time quantitative RT-PCR using specific primers for Ki67 ( C ), Hmox1 , Sod1 , Gclc , Gclm , Gsta1 , and Gapdh ( D ). Values are the amount of each mRNA relative to that of Gapdh and represent the means ± S.D. for three independent experiments. Statistically significant differences are indicated (**, p

    Article Snippet: Real-time quantitative PCR was performed using the One-Step SYBR PrimeScript RT-PCR Kit II (Takara Bio) on a CFX96 Touch real-time PCR detection system (Bio-Rad) according to the manufacturer's manual.

    Techniques: Mouse Assay, SYBR Green Assay, Quantitative RT-PCR

    Expression profiles of Aldh3 family members in various tissues. Total RNAs prepared from liver, kidney, retina, cornea, brain, small intestine, lung, testis, spleen, dermis, and epidermis tissue ( A and C ) and from keratinocytes kept undifferentiated or differentiated for 4 days ( B and D ) obtained from wild-type ( A–D ) and Aldh3a2 KO mice ( C and D ) were subjected to SYBR Green-based real-time quantitative RT-PCR using specific primers for Aldh3a1 , Aldh3a2 , Aldh3b1 , Aldh3b2 , Aldh3b3 , and Gapdh . Values are the amount of each mRNA relative to that of Gapdh , and represent the means ± S.D. for three independent experiments. A statistically significant difference is indicated (**, p

    Journal: The Journal of Biological Chemistry

    Article Title: Disruption of the Sjögren-Larsson Syndrome Gene Aldh3a2 in Mice Increases Keratinocyte Growth and Retards Skin Barrier Recovery *

    doi: 10.1074/jbc.M116.714030

    Figure Lengend Snippet: Expression profiles of Aldh3 family members in various tissues. Total RNAs prepared from liver, kidney, retina, cornea, brain, small intestine, lung, testis, spleen, dermis, and epidermis tissue ( A and C ) and from keratinocytes kept undifferentiated or differentiated for 4 days ( B and D ) obtained from wild-type ( A–D ) and Aldh3a2 KO mice ( C and D ) were subjected to SYBR Green-based real-time quantitative RT-PCR using specific primers for Aldh3a1 , Aldh3a2 , Aldh3b1 , Aldh3b2 , Aldh3b3 , and Gapdh . Values are the amount of each mRNA relative to that of Gapdh , and represent the means ± S.D. for three independent experiments. A statistically significant difference is indicated (**, p

    Article Snippet: Real-time quantitative PCR was performed using the One-Step SYBR PrimeScript RT-PCR Kit II (Takara Bio) on a CFX96 Touch real-time PCR detection system (Bio-Rad) according to the manufacturer's manual.

    Techniques: Expressing, Mouse Assay, SYBR Green Assay, Quantitative RT-PCR

    Melting peaks of the SYBR Green Ⅰ-based one-step qRT-PCR products. ( A ) Melting peaks of the qRT-PCR products obtained from in vitro transcribed HTNV cRNA ranging from 1×10 8 to 1×10 3 copies/µl. Only a single sharp peak at 84°C was visible in the melt peak chart. ( B ) Melting peaks obtained from serum samples of HFRS patients. All melting curves in A and B have similar shapes.

    Journal: PLoS ONE

    Article Title: Quantification of Hantaan Virus with a SYBR Green I-Based One-Step qRT-PCR Assay

    doi: 10.1371/journal.pone.0081525

    Figure Lengend Snippet: Melting peaks of the SYBR Green Ⅰ-based one-step qRT-PCR products. ( A ) Melting peaks of the qRT-PCR products obtained from in vitro transcribed HTNV cRNA ranging from 1×10 8 to 1×10 3 copies/µl. Only a single sharp peak at 84°C was visible in the melt peak chart. ( B ) Melting peaks obtained from serum samples of HFRS patients. All melting curves in A and B have similar shapes.

    Article Snippet: SYBR Green Ⅰ-based one-step qRT-PCR assay qRT-PCR amplification was performed using a Bio-Rad iQ5 Multi-color Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) with the One-Step SYBR PrimeScript RT-PCR Kit ii (TaKaRa Biotechnology, Dalian, China).

    Techniques: SYBR Green Assay, Quantitative RT-PCR, In Vitro

    Sensitivity of the SYBR Green Ⅰ-based one-step qRT-PCR assay. ( A ) Serial dilutions of the HTNV cRNA standards ranging from 1×10 6 to 1×10 -1 copies/µl were amplified using the SYBR Green Ⅰ-based one-step qRT-PCR assay in duplicate, and the minimum detection limit of the qRT-PCR assay was 10 copies/µl. ( B ) The qRT-PCR products were electrophoresed on a 2% agarose gel. Marker: DNA ladder of DL2000.

    Journal: PLoS ONE

    Article Title: Quantification of Hantaan Virus with a SYBR Green I-Based One-Step qRT-PCR Assay

    doi: 10.1371/journal.pone.0081525

    Figure Lengend Snippet: Sensitivity of the SYBR Green Ⅰ-based one-step qRT-PCR assay. ( A ) Serial dilutions of the HTNV cRNA standards ranging from 1×10 6 to 1×10 -1 copies/µl were amplified using the SYBR Green Ⅰ-based one-step qRT-PCR assay in duplicate, and the minimum detection limit of the qRT-PCR assay was 10 copies/µl. ( B ) The qRT-PCR products were electrophoresed on a 2% agarose gel. Marker: DNA ladder of DL2000.

    Article Snippet: SYBR Green Ⅰ-based one-step qRT-PCR assay qRT-PCR amplification was performed using a Bio-Rad iQ5 Multi-color Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) with the One-Step SYBR PrimeScript RT-PCR Kit ii (TaKaRa Biotechnology, Dalian, China).

    Techniques: SYBR Green Assay, Quantitative RT-PCR, Amplification, Agarose Gel Electrophoresis, Marker

    Amplification chart and melt peak chart of HTNV and SEOV. ( A ) Different dilutions of HTNV and SEOV RNA were tested using the SYBR Green Ⅰ-based one-step qRT-PCR assay, and only HTNV RNA was amplified. ( B ) A single sharp peak at 84°C in the melt peak chart indicated that only specific products of HTNV was obtained.

    Journal: PLoS ONE

    Article Title: Quantification of Hantaan Virus with a SYBR Green I-Based One-Step qRT-PCR Assay

    doi: 10.1371/journal.pone.0081525

    Figure Lengend Snippet: Amplification chart and melt peak chart of HTNV and SEOV. ( A ) Different dilutions of HTNV and SEOV RNA were tested using the SYBR Green Ⅰ-based one-step qRT-PCR assay, and only HTNV RNA was amplified. ( B ) A single sharp peak at 84°C in the melt peak chart indicated that only specific products of HTNV was obtained.

    Article Snippet: SYBR Green Ⅰ-based one-step qRT-PCR assay qRT-PCR amplification was performed using a Bio-Rad iQ5 Multi-color Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) with the One-Step SYBR PrimeScript RT-PCR Kit ii (TaKaRa Biotechnology, Dalian, China).

    Techniques: Amplification, SYBR Green Assay, Quantitative RT-PCR

    Effects of Lico A exposure on the upregulation of Nrf2-mediated antioxidant signaling pathway in HepG2 cells. (A) HepG2 cells were treated with different concentrations of Lico A (0.4625, 0.925, 1.85, or 3.7 μM) for 18 h, and (B) cells were exposed to Lico A (3.7 μM) for four time points (1, 3, 6, or 18 h). Protein expressions of GCLC, GCLM, HO-1, and NQO1 were measure by Western blotting analysis. (C,D) Quantification of GCLC, GCLM, HO-1, and NQO1 protein expressions were performed by densitometric analysis and β-actin was acted as an internal control. (E) Cells were exposed to Lico A (3.7 μM) for three time points (3, 6, or 18 h). Effects of Lico A on GCLC, GCLM, HO-1, and NQO1 genes expression. Total RNA was extracted from HepG2 cells and genes expression was quantified using real-time PCR. Similar results were obtained from three independent experiments. All results were expressed as means ± SEM of three independent experiments. # p

    Journal: Frontiers in Pharmacology

    Article Title: Licochalcone A Upregulates Nrf2 Antioxidant Pathway and Thereby Alleviates Acetaminophen-Induced Hepatotoxicity

    doi: 10.3389/fphar.2018.00147

    Figure Lengend Snippet: Effects of Lico A exposure on the upregulation of Nrf2-mediated antioxidant signaling pathway in HepG2 cells. (A) HepG2 cells were treated with different concentrations of Lico A (0.4625, 0.925, 1.85, or 3.7 μM) for 18 h, and (B) cells were exposed to Lico A (3.7 μM) for four time points (1, 3, 6, or 18 h). Protein expressions of GCLC, GCLM, HO-1, and NQO1 were measure by Western blotting analysis. (C,D) Quantification of GCLC, GCLM, HO-1, and NQO1 protein expressions were performed by densitometric analysis and β-actin was acted as an internal control. (E) Cells were exposed to Lico A (3.7 μM) for three time points (3, 6, or 18 h). Effects of Lico A on GCLC, GCLM, HO-1, and NQO1 genes expression. Total RNA was extracted from HepG2 cells and genes expression was quantified using real-time PCR. Similar results were obtained from three independent experiments. All results were expressed as means ± SEM of three independent experiments. # p

    Article Snippet: After the concentration of RNA was determined by spectrophotometer, 1 μg of RNA was transformed into cDNA using Prime-Script RT-PCR kit (Takara).

    Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction

    LncATB functions by binding to miR-200 family members. a RNA pull-down followed by microRNA qRT-PCR assays to detect miR-200 family members endogenously associated with lncATB. b MCF-7 cell lysates were incubated with biotin-labelled lncATB; after the pull-down, miR-200 miRNAs were extracted and assessed by qRT-PCR assays. c − g Relative luciferase activity in MCF-7 cells transfected with specific miR-200 luciferase reporters in the presence of pcDNA3.1-ATB alone or with the cotransfection of c miR-200a mimics, d miR-200b mimics, e miR-200c mimics, f miR-141 mimics, or g miR-429 mimics. h − l Relative luciferase activity in BT-549 cells cotransfected with specific miR-200 luciferase reporters and shATB-#1/2. For each specific reporter, h miR-200a inhibitor, i miR-200b inhibitor, j miR-200c inhibitor, k miR-141 inhibitor, or l miR-429 inhibitor was also added. * P

    Journal: Cell Death & Disease

    Article Title: Long noncoding RNA ATB promotes the epithelial−mesenchymal transition by upregulating the miR-200c/Twist1 axe and predicts poor prognosis in breast cancer

    doi: 10.1038/s41419-018-1210-9

    Figure Lengend Snippet: LncATB functions by binding to miR-200 family members. a RNA pull-down followed by microRNA qRT-PCR assays to detect miR-200 family members endogenously associated with lncATB. b MCF-7 cell lysates were incubated with biotin-labelled lncATB; after the pull-down, miR-200 miRNAs were extracted and assessed by qRT-PCR assays. c − g Relative luciferase activity in MCF-7 cells transfected with specific miR-200 luciferase reporters in the presence of pcDNA3.1-ATB alone or with the cotransfection of c miR-200a mimics, d miR-200b mimics, e miR-200c mimics, f miR-141 mimics, or g miR-429 mimics. h − l Relative luciferase activity in BT-549 cells cotransfected with specific miR-200 luciferase reporters and shATB-#1/2. For each specific reporter, h miR-200a inhibitor, i miR-200b inhibitor, j miR-200c inhibitor, k miR-141 inhibitor, or l miR-429 inhibitor was also added. * P

    Article Snippet: The RNA was reverse-transcribed into cDNA using the Prime-Script RT-PCR kit (Takara, Dalian, China) following the manufacturer’s instructions.

    Techniques: Binding Assay, Quantitative RT-PCR, Incubation, Luciferase, Activity Assay, Transfection, Cotransfection