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tiangen biotech co real time pcr total rna
The interaction between EIF3B and piR-823 in LX-2 cells. ( A ) We collected the <t>RNA-protein</t> solution from RNA pull-down experiment and Coomassie brilliant blue staining showed that the piR-823 channel had 1 additional protein band as compared to the control channel (red box, indicated by black arrow). ( B ) Western blot analysis to test the EIF3B combined with piR-823. ( C ) Real-time <t>PCR</t> to test the piR-823 RNA combined with EIF3B. Bars=means ±SEM. n=3, * P
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1) Product Images from "The Combination of piR-823 and Eukaryotic Initiation Factor 3 B (EIF3B) Activates Hepatic Stellate Cells via Upregulating TGF-β1 in Liver Fibrogenesis"

Article Title: The Combination of piR-823 and Eukaryotic Initiation Factor 3 B (EIF3B) Activates Hepatic Stellate Cells via Upregulating TGF-β1 in Liver Fibrogenesis

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

doi: 10.12659/MSM.914222

The interaction between EIF3B and piR-823 in LX-2 cells. ( A ) We collected the RNA-protein solution from RNA pull-down experiment and Coomassie brilliant blue staining showed that the piR-823 channel had 1 additional protein band as compared to the control channel (red box, indicated by black arrow). ( B ) Western blot analysis to test the EIF3B combined with piR-823. ( C ) Real-time PCR to test the piR-823 RNA combined with EIF3B. Bars=means ±SEM. n=3, * P
Figure Legend Snippet: The interaction between EIF3B and piR-823 in LX-2 cells. ( A ) We collected the RNA-protein solution from RNA pull-down experiment and Coomassie brilliant blue staining showed that the piR-823 channel had 1 additional protein band as compared to the control channel (red box, indicated by black arrow). ( B ) Western blot analysis to test the EIF3B combined with piR-823. ( C ) Real-time PCR to test the piR-823 RNA combined with EIF3B. Bars=means ±SEM. n=3, * P

Techniques Used: Staining, Western Blot, Real-time Polymerase Chain Reaction

2) Product Images from "shRNA-Mediated Silencing of Y-Box Binding Protein-1 (YB-1) Suppresses Growth of Neuroblastoma Cell SH-SY5Y In Vitro and In Vivo"

Article Title: shRNA-Mediated Silencing of Y-Box Binding Protein-1 (YB-1) Suppresses Growth of Neuroblastoma Cell SH-SY5Y In Vitro and In Vivo

Journal: PLoS ONE

doi: 10.1371/journal.pone.0127224

YB-1 regulated Cyclin D1 transcription in neuroblastoma SH-SY5Y cells. (A) Expression levels of cell cycle regulators such as Cyclin A and Cyclin D1 in SH-SY5Y, SH-shCON and SH-shYB-1 cells were examined by Western blot analysis. (B) A schematic illustration of the promoter region of CCND1 which encodes Cyclin D1 indicates the transcription start site (TSS), putative YB-1 binding site and the location of two sets of primers (CCND1p #1 and #2) used for chromatin-immunoprecipitation (ChIP) assay. (C) ChIP assay followed by PCR analysis was performed in SH-SY5Y cells to detect binding of YB-1 on the promoter region of CCND1. Binding of RNA polymerase II (RNA PolII) on GAPDH promoter, which was detected with primers for GAPDH promoter (GAPDHp) by PCR, was used as a positive control for ChIP experiments, whereas IgG served as a negative control for non-specific binding. (D) pGL-3 Firefly luciferase reporter plasmid containing a CCND1 promoter fragment (pGL-3-CCND1) or pGL-3 vector alone was transfected in combination with Renilla luciferase reporter vector pRL-TK into SH-SY5Y, SH-shCON and SH-shYB-1 cells. Luciferase activity representing activity of the promoter was quantified as the ratio of FL/RL which was then normalized to SH-SY5Y cells. Values are expressed as mean ± standard deviation. Compared with SH-SY5Y control, *** P
Figure Legend Snippet: YB-1 regulated Cyclin D1 transcription in neuroblastoma SH-SY5Y cells. (A) Expression levels of cell cycle regulators such as Cyclin A and Cyclin D1 in SH-SY5Y, SH-shCON and SH-shYB-1 cells were examined by Western blot analysis. (B) A schematic illustration of the promoter region of CCND1 which encodes Cyclin D1 indicates the transcription start site (TSS), putative YB-1 binding site and the location of two sets of primers (CCND1p #1 and #2) used for chromatin-immunoprecipitation (ChIP) assay. (C) ChIP assay followed by PCR analysis was performed in SH-SY5Y cells to detect binding of YB-1 on the promoter region of CCND1. Binding of RNA polymerase II (RNA PolII) on GAPDH promoter, which was detected with primers for GAPDH promoter (GAPDHp) by PCR, was used as a positive control for ChIP experiments, whereas IgG served as a negative control for non-specific binding. (D) pGL-3 Firefly luciferase reporter plasmid containing a CCND1 promoter fragment (pGL-3-CCND1) or pGL-3 vector alone was transfected in combination with Renilla luciferase reporter vector pRL-TK into SH-SY5Y, SH-shCON and SH-shYB-1 cells. Luciferase activity representing activity of the promoter was quantified as the ratio of FL/RL which was then normalized to SH-SY5Y cells. Values are expressed as mean ± standard deviation. Compared with SH-SY5Y control, *** P

Techniques Used: Expressing, Western Blot, Binding Assay, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Positive Control, Negative Control, Luciferase, Plasmid Preparation, Transfection, Activity Assay, Standard Deviation

3) Product Images from "Chromosome doubling mediates superior drought tolerance in Lycium ruthenicum via abscisic acid signaling"

Article Title: Chromosome doubling mediates superior drought tolerance in Lycium ruthenicum via abscisic acid signaling

Journal: Horticulture Research

doi: 10.1038/s41438-020-0260-1

Expression patterns of 12 candidate genes according to RNA-seq (white) and qRT-PCR (oblique line) for selected transcripts. The data represent the mean ± SD of three independent experiments. The X -axis shows the selected gene ID, and the Y -axis shows the log 2 ratio
Figure Legend Snippet: Expression patterns of 12 candidate genes according to RNA-seq (white) and qRT-PCR (oblique line) for selected transcripts. The data represent the mean ± SD of three independent experiments. The X -axis shows the selected gene ID, and the Y -axis shows the log 2 ratio

Techniques Used: Expressing, RNA Sequencing Assay, Quantitative RT-PCR

4) Product Images from "Inhibition of A20 expression in tumor microenvironment exerts anti-tumor effect through inducing myeloid-derived suppressor cells apoptosis"

Article Title: Inhibition of A20 expression in tumor microenvironment exerts anti-tumor effect through inducing myeloid-derived suppressor cells apoptosis

Journal: Scientific Reports

doi: 10.1038/srep16437

A20 expression in tumor microenvironment and the knockdown of A20 by si-RNA. ( a ) The expression of A20 in different cell lines. A20 expression was tested by flow cytometry in E.G7, B16-F10, CT26, 4T1 and L929 cell lines. Total number of 30000 cells was analyzed. The grey area represents the isotype control. si-A20 was used to test the specificity of the staining. The efficiency of knockdown of A20 was illustrated as reduction in mean fluorescence intensity. ( b ) Representative IHC analysis of A20 in E.G7 tumor sections ( n = 5). Scale bar 50 μm. ( c ) The immunofluorescence staining of A20-positive and Gr1-positive cells. E.G7 tumor frozen sections were stained with DAPI (blue), Gr1 (Alexa-595) and A20 (FITC) ( n = 5). Scale bar 50μm. ( d,e ) Knockdown of A20 by si-RNA in L929 cell lines was confirmed by qRT-PCR and western blotting. L929 cells were transfected with si-RNA in vitro and the mRNA level and protein level of A20 were detected. ( f ) Decrease of A20 expression in tumor tissue after si-RNA treatment. E.G7 tumor-bearing mice were treated with si-RNA intratumorally and CD11b + cells were isolated from pooled tumor tissue suspension (n = 3–5). Data were obtained from two independent experiments. Data represent means ± SD. * p
Figure Legend Snippet: A20 expression in tumor microenvironment and the knockdown of A20 by si-RNA. ( a ) The expression of A20 in different cell lines. A20 expression was tested by flow cytometry in E.G7, B16-F10, CT26, 4T1 and L929 cell lines. Total number of 30000 cells was analyzed. The grey area represents the isotype control. si-A20 was used to test the specificity of the staining. The efficiency of knockdown of A20 was illustrated as reduction in mean fluorescence intensity. ( b ) Representative IHC analysis of A20 in E.G7 tumor sections ( n = 5). Scale bar 50 μm. ( c ) The immunofluorescence staining of A20-positive and Gr1-positive cells. E.G7 tumor frozen sections were stained with DAPI (blue), Gr1 (Alexa-595) and A20 (FITC) ( n = 5). Scale bar 50μm. ( d,e ) Knockdown of A20 by si-RNA in L929 cell lines was confirmed by qRT-PCR and western blotting. L929 cells were transfected with si-RNA in vitro and the mRNA level and protein level of A20 were detected. ( f ) Decrease of A20 expression in tumor tissue after si-RNA treatment. E.G7 tumor-bearing mice were treated with si-RNA intratumorally and CD11b + cells were isolated from pooled tumor tissue suspension (n = 3–5). Data were obtained from two independent experiments. Data represent means ± SD. * p

Techniques Used: Expressing, Flow Cytometry, Cytometry, Staining, Fluorescence, Immunohistochemistry, Immunofluorescence, Quantitative RT-PCR, Western Blot, Transfection, In Vitro, Mouse Assay, Isolation

5) Product Images from "Association between functional polymorphisms in IL‐33/ST2 pathway and risk of osteosarcoma. Association between functional polymorphisms in IL‐33/ST2 pathway and risk of osteosarcoma"

Article Title: Association between functional polymorphisms in IL‐33/ST2 pathway and risk of osteosarcoma. Association between functional polymorphisms in IL‐33/ST2 pathway and risk of osteosarcoma

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13653

The rs3821204 G/C influences the transcriptional activity and ST 2 mRNA expression by altering the binding site of miR‐202‐3p. A, In silico prediction of miR‐203‐3p binding site to ST 2 3′ UTR containing rs3821204 G or C allele. B, The rs3821204 G or C plasmid was cotransfected with miR‐202‐3p mimics or negative control into HEK 293 cells. Relative luciferase activity was measured using the dual luciferase reporter assay. C, mi RNA ‐202‐3p mimics or negative control was transfected into Saos‐2 (rs3821204 GG ) and MG ‐63 (rs3821204 CC ) cells. Relative expression of ST 2 mRNA was analysed using real‐time PCR . Data were expressed by mean ± standard error (* P
Figure Legend Snippet: The rs3821204 G/C influences the transcriptional activity and ST 2 mRNA expression by altering the binding site of miR‐202‐3p. A, In silico prediction of miR‐203‐3p binding site to ST 2 3′ UTR containing rs3821204 G or C allele. B, The rs3821204 G or C plasmid was cotransfected with miR‐202‐3p mimics or negative control into HEK 293 cells. Relative luciferase activity was measured using the dual luciferase reporter assay. C, mi RNA ‐202‐3p mimics or negative control was transfected into Saos‐2 (rs3821204 GG ) and MG ‐63 (rs3821204 CC ) cells. Relative expression of ST 2 mRNA was analysed using real‐time PCR . Data were expressed by mean ± standard error (* P

Techniques Used: Activity Assay, Expressing, Binding Assay, In Silico, Plasmid Preparation, Negative Control, Luciferase, Reporter Assay, Transfection, Real-time Polymerase Chain Reaction

6) Product Images from "piR‐823 contributes to colorectal tumorigenesis by enhancing the transcriptional activity of HSF1"

Article Title: piR‐823 contributes to colorectal tumorigenesis by enhancing the transcriptional activity of HSF1

Journal: Cancer Science

doi: 10.1111/cas.13300

piR‐823 regulates HSF 1 activity by binding to HSF 1 and modulating its phosphorylation at Ser 326 . (a, b) mRNA levels (a) and protein levels (b) of HSF 1 was evaluated in HCT 116 cells transfected with Ant‐823 or NC . (c) The transcriptional activity of HSF 1 was assessed by dual‐luciferase reporter assay in HCT 116 cells. (d, e) Nuclear translocation of HSF 1 in HCT 116 cells was evaluated by western blot analysis of cytoplasmic and nuclear extractions (d) and immunofluorescence staining for HSF 1 (green) (e). Nuclear DNA was stained with DAPI (blue). Scale bar, 20 μm. (f) The expression of p S er326 HSF 1 in HCT 116 cells was assessed by western blot analysis. (d) Cellular localization of p S er326 HSF 1 in HCT 116 cells was determined by western blot analysis. (h) The interaction of piR‐823 with HSF 1 in HCT 116 cells was assessed by RIP assay using anti‐ HSF 1 or IgG antibody as a control. The eluted RNA was analyzed by real‐time PCR . Immunoprecipitation efficiency was assessed by western blot analysis. Data are shown as the mean ± SD from three independent experiments, ** P
Figure Legend Snippet: piR‐823 regulates HSF 1 activity by binding to HSF 1 and modulating its phosphorylation at Ser 326 . (a, b) mRNA levels (a) and protein levels (b) of HSF 1 was evaluated in HCT 116 cells transfected with Ant‐823 or NC . (c) The transcriptional activity of HSF 1 was assessed by dual‐luciferase reporter assay in HCT 116 cells. (d, e) Nuclear translocation of HSF 1 in HCT 116 cells was evaluated by western blot analysis of cytoplasmic and nuclear extractions (d) and immunofluorescence staining for HSF 1 (green) (e). Nuclear DNA was stained with DAPI (blue). Scale bar, 20 μm. (f) The expression of p S er326 HSF 1 in HCT 116 cells was assessed by western blot analysis. (d) Cellular localization of p S er326 HSF 1 in HCT 116 cells was determined by western blot analysis. (h) The interaction of piR‐823 with HSF 1 in HCT 116 cells was assessed by RIP assay using anti‐ HSF 1 or IgG antibody as a control. The eluted RNA was analyzed by real‐time PCR . Immunoprecipitation efficiency was assessed by western blot analysis. Data are shown as the mean ± SD from three independent experiments, ** P

Techniques Used: Activity Assay, Binding Assay, Transfection, Luciferase, Reporter Assay, Translocation Assay, Western Blot, Immunofluorescence, Staining, Expressing, Real-time Polymerase Chain Reaction, Immunoprecipitation

7) Product Images from "ITE and TCDD Differentially Regulate the Vascular Remodeling of Rat Placenta via the Activation of AhR"

Article Title: ITE and TCDD Differentially Regulate the Vascular Remodeling of Rat Placenta via the Activation of AhR

Journal: PLoS ONE

doi: 10.1371/journal.pone.0086549

Effects of ITE and TCDD on mRNA expression of angiogenic-associated genes in placental tissues. The total RNA samples from placental tissues were subjected to real-time PCR. *Differ from the control ( p
Figure Legend Snippet: Effects of ITE and TCDD on mRNA expression of angiogenic-associated genes in placental tissues. The total RNA samples from placental tissues were subjected to real-time PCR. *Differ from the control ( p

Techniques Used: Expressing, Real-time Polymerase Chain Reaction

8) Product Images from "Effect of gap junctions on RAW264.7 macrophages infected with H37Rv"

Article Title: Effect of gap junctions on RAW264.7 macrophages infected with H37Rv

Journal: Medicine

doi: 10.1097/MD.0000000000012125

H37Rv infection induces host macrophage connexin expression. RAW264.7 cells were infected by H37Rv strains for 12 hours. Total RNA and protein were tested using RT-PCR (Cx37 and Cx43 mRNA) (A) and Western blotting (Cx43 protein) (B). Both the control and infection cells were collected and tested using immunofluorescence analysis (C). Data are expressed as mean±SD from 3 independent experiments. ∗ P
Figure Legend Snippet: H37Rv infection induces host macrophage connexin expression. RAW264.7 cells were infected by H37Rv strains for 12 hours. Total RNA and protein were tested using RT-PCR (Cx37 and Cx43 mRNA) (A) and Western blotting (Cx43 protein) (B). Both the control and infection cells were collected and tested using immunofluorescence analysis (C). Data are expressed as mean±SD from 3 independent experiments. ∗ P

Techniques Used: Infection, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Immunofluorescence

9) Product Images from "MiR-375 attenuates injury of cerebral ischemia/reperfusion via targetting Ctgf"

Article Title: MiR-375 attenuates injury of cerebral ischemia/reperfusion via targetting Ctgf

Journal: Bioscience Reports

doi: 10.1042/BSR20171242

. MiR-375 attenuated infarction volumes in I/R rat brain ( A ) TTC staining of representative coronal sections after I/R. The relative infarct area percentage was evaluated by observing the unstained infarcted tissue zone (white) and the stained normal tissue zone (red). ( B ) The protein expression of Ctgf in Sham rat brains and I/R rat brains treated with NC, mimic, and FNS was analyzed by Western blotting. ( C , D ) RNA expression of miR-375 (C) and Ctgf (D) in Sham rat brains and I/R rat brains treated with NC, mimic, and FNS was analyzed by qRT-PCR. ( E ) The protein expression of Ctgf in sham rat brains and I/R rat brains treated with NC, mimics and FNS was analyzed by western blotting. The experiments were performed in triplicate and each value represented mean ± S.D. *** P
Figure Legend Snippet: . MiR-375 attenuated infarction volumes in I/R rat brain ( A ) TTC staining of representative coronal sections after I/R. The relative infarct area percentage was evaluated by observing the unstained infarcted tissue zone (white) and the stained normal tissue zone (red). ( B ) The protein expression of Ctgf in Sham rat brains and I/R rat brains treated with NC, mimic, and FNS was analyzed by Western blotting. ( C , D ) RNA expression of miR-375 (C) and Ctgf (D) in Sham rat brains and I/R rat brains treated with NC, mimic, and FNS was analyzed by qRT-PCR. ( E ) The protein expression of Ctgf in sham rat brains and I/R rat brains treated with NC, mimics and FNS was analyzed by western blotting. The experiments were performed in triplicate and each value represented mean ± S.D. *** P

Techniques Used: Staining, Expressing, Western Blot, RNA Expression, Quantitative RT-PCR

MiR-375 promoted the proliferation and migration, and inhibited the apoptosis in H/R PC12 cells ( A , B ) The RNA expressions of miR-375 and Ctgf (A) and the protein expression of Ctgf (B) in PC12 cells subjected to H/R or not. ( C ) The RNA expressions of miR-375 and Ctgf in H/R PC12 cells treated with NC or miR-375 . The expression of RNA was determined by qRT-PCR, and the expression of protein was determined by Western blotting. ( D ) The effect of miR-375 on H/R p12 cells proliferation was detected by CCK-8 assay. ( E ) The effect of miR-375 on H/R p12 cells apoptosis was examined by Hoechst 33258 staining. ( F ) The expressions of Ctgf in H/R PC12 cells treated with NC or miR-375 was detected by immunofluorescence staining. ( G , H ) The effect of miR-375 on H/R p12 cell migration was detected by transwell assay, and the migrant cells were calculated. ( I ) The effect of miR-375 on protein expression of Ctgf, p21, PI3K, and p-AKT in H/R p12 cells was examined by Western blotting. The experiments were performed in triplicate and each value represents mean ± S.D. *** P
Figure Legend Snippet: MiR-375 promoted the proliferation and migration, and inhibited the apoptosis in H/R PC12 cells ( A , B ) The RNA expressions of miR-375 and Ctgf (A) and the protein expression of Ctgf (B) in PC12 cells subjected to H/R or not. ( C ) The RNA expressions of miR-375 and Ctgf in H/R PC12 cells treated with NC or miR-375 . The expression of RNA was determined by qRT-PCR, and the expression of protein was determined by Western blotting. ( D ) The effect of miR-375 on H/R p12 cells proliferation was detected by CCK-8 assay. ( E ) The effect of miR-375 on H/R p12 cells apoptosis was examined by Hoechst 33258 staining. ( F ) The expressions of Ctgf in H/R PC12 cells treated with NC or miR-375 was detected by immunofluorescence staining. ( G , H ) The effect of miR-375 on H/R p12 cell migration was detected by transwell assay, and the migrant cells were calculated. ( I ) The effect of miR-375 on protein expression of Ctgf, p21, PI3K, and p-AKT in H/R p12 cells was examined by Western blotting. The experiments were performed in triplicate and each value represents mean ± S.D. *** P

Techniques Used: Migration, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Staining, Immunofluorescence, Transwell Assay

10) Product Images from "Ancestor of land plants acquired the DNA-3-methyladenine glycosylase (MAG) gene from bacteria through horizontal gene transfer"

Article Title: Ancestor of land plants acquired the DNA-3-methyladenine glycosylase (MAG) gene from bacteria through horizontal gene transfer

Journal: Scientific Reports

doi: 10.1038/s41598-017-05066-w

Real-time PCR analysis for the expression of ZmMAG in response to UV and zeocin treatments. Two-week-old maize seedlings grown in soil were collected for gene expression analysis of different time intervals. Total RNA was prepared from 2-week-old seedlings of wild-type maize after UV and zeocin treatments, respectively, and then reverse-transcribed. The resultant cDNAs were used as templates for real-time PCR analysis and ZmActin was used as an internal control. Real-time PCR was performed with ZmMAG specific primers and ZmActin specific primers. Data represent means and standard deviation of three replicates. Different letters following mean values indicate significant difference ( P
Figure Legend Snippet: Real-time PCR analysis for the expression of ZmMAG in response to UV and zeocin treatments. Two-week-old maize seedlings grown in soil were collected for gene expression analysis of different time intervals. Total RNA was prepared from 2-week-old seedlings of wild-type maize after UV and zeocin treatments, respectively, and then reverse-transcribed. The resultant cDNAs were used as templates for real-time PCR analysis and ZmActin was used as an internal control. Real-time PCR was performed with ZmMAG specific primers and ZmActin specific primers. Data represent means and standard deviation of three replicates. Different letters following mean values indicate significant difference ( P

Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Standard Deviation

11) Product Images from "Deficiency of PRKD2 triggers hyperinsulinemia and metabolic disorders"

Article Title: Deficiency of PRKD2 triggers hyperinsulinemia and metabolic disorders

Journal: Nature Communications

doi: 10.1038/s41467-018-04352-z

PRKD2 K410X variation is shared by two hyperinsulinemic monkeys. a Sanger sequencing indicating one heterozygous nonsense variation in PRKD2 (K410X) exclusive to the two cases (950807, 960109). b As for the PRKD2 nonsense variation, the allele frequencies of the two alleles (WT, Mutant) were shown, according to the RNA-seq data in muscle and adipose tissue from a PRKD2 mutant monkey (950807). c Real-time PCR quantifications of the PRKD2 expression in skeletal muscle of WT and the PRKD2 mutant monkeys. d Western blots of PRKD 2 protein expression in skeletal muscle (left) and adipose tissues (right) from control (black) and hyperinsulinemic (red) monkeys
Figure Legend Snippet: PRKD2 K410X variation is shared by two hyperinsulinemic monkeys. a Sanger sequencing indicating one heterozygous nonsense variation in PRKD2 (K410X) exclusive to the two cases (950807, 960109). b As for the PRKD2 nonsense variation, the allele frequencies of the two alleles (WT, Mutant) were shown, according to the RNA-seq data in muscle and adipose tissue from a PRKD2 mutant monkey (950807). c Real-time PCR quantifications of the PRKD2 expression in skeletal muscle of WT and the PRKD2 mutant monkeys. d Western blots of PRKD 2 protein expression in skeletal muscle (left) and adipose tissues (right) from control (black) and hyperinsulinemic (red) monkeys

Techniques Used: Sequencing, Mutagenesis, RNA Sequencing Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot

Related Articles

Real-time Polymerase Chain Reaction:

Article Title: Chromosome doubling mediates superior drought tolerance in Lycium ruthenicum via abscisic acid signaling
Article Snippet: .. Validation of DEGs by quantitative real-time PCR Total RNA was extracted using a plant polysaccharide polyphenol RNA kit (TIANGEN Biotech Co., Ltd, Beijing, China). .. For each sample, 1 ng of RNA was used as the template for reverse transcription to obtain the same concentration of cDNA using a reverse transcription kit (Aidlab Biotechnologies Co., Ltd, Beijing, China).

Article Title: Plastid-nucleus communication involves calcium-modulated MAPK signalling
Article Snippet: .. RNA extraction and quantitative real-time PCR Total RNA was extracted from seedlings by RNA extraction kit (Tiangen). .. After DNase I treatment, reverse transcription was conducted using PrimeScript II 1st Strand cDNA Synthesis Kit (Takara) according to the manufacturer's protocol.

Article Title: The Combination of piR-823 and Eukaryotic Initiation Factor 3 B (EIF3B) Activates Hepatic Stellate Cells via Upregulating TGF-β1 in Liver Fibrogenesis
Article Snippet: .. Isolation of RNA and real-time PCR Total RNA was extracted using a Pure Cell/Bacteria kit (Tiangen, Beijing, China). .. The miScript Plant RT Kit (Qiagen, Hilden, Germany) designed for small RNAs 3′end with 2′-O-Me modification was used for small RNA reverse transcription.

Article Title: Inhibition of A20 expression in tumor microenvironment exerts anti-tumor effect through inducing myeloid-derived suppressor cells apoptosis
Article Snippet: .. Real-Time PCR Total RNA was prepared using the RNA Simple Total RNA Kit (TIANGEN), and the isolated RNA was reverse transcribed into cDNA using a PrimeScript™ RT reagent Kit with gDNA Eraser (TaKaRa). .. Primers used were as follows: A20 forward 5′-TTT GCT ACG ACA CTC GGA AC-3′ and reverse 5′-TTC TGA GGA TGT TGC TGA GG-3′; GAPDH forward 5′-CCC AGA AGA CTG TGG ATG G-3′ and reverse 5′-CAC ATT GGG GGT AGG AAC AC-3′.

Article Title: shRNA-Mediated Silencing of Y-Box Binding Protein-1 (YB-1) Suppresses Growth of Neuroblastoma Cell SH-SY5Y In Vitro and In Vivo
Article Snippet: .. RNA isolation and real-time PCR Total RNA of SH-SY5Y parental cells, YB-1-silenced SH-SY5Y cells (SH-shYB-1) and SH-shCON cells was isolated with RNAsimple Total RNA Kit (TIANGEN Biotech, Beijing, China), followed by reverse transcription using Super M-MLV reverse transcriptase (BioTeke, Beijing, China). .. Real-time PCR was performed to determine the levels of YB-1 and internal control β-actin using SYBR Green Master Mix (Solarbio, Beijing, China) and respective primers in an Exicycler 96 quantitative fluorescence analyzer (Bioneer, Daejeon, Korea).

Article Title: SiMYB56 Confers Drought Stress Tolerance in Transgenic Rice by Regulating Lignin Biosynthesis and ABA Signaling Pathway
Article Snippet: .. RNA Isolation and Quantitative Real-Time PCR Total RNA was extracted from seedlings using the Total RNA Extraction Kit (TIANGEN, China). .. The cDNA was synthesized in accordance with Fast Quant RT Super Mix Reverse Transcription Kit instructions (TransGene, Beijing, China).

Article Title: Inhibiting Receptor of Advanced Glycation End Products Attenuates Pressure Overload-Induced Cardiac Dysfunction by Preventing Excessive Autophagy
Article Snippet: .. Quantitative Real-Time PCR Total RNA was extracted from LV tissue by using an RNAprep Pure Tissue Kit (Tiangen, Beijing, China), after which first-strand cDNA was synthesized with a FastKing RT Kit (Tiangen, Beijing, China) and quantitative PCR (qPCR) was performed using SYBR® Premix Ex TaqTM II (Takara, Beijing, China). .. Target gene expression was normalized for that of GAPDH and compared among the groups.

Article Title: Association between functional polymorphisms in IL‐33/ST2 pathway and risk of osteosarcoma. Association between functional polymorphisms in IL‐33/ST2 pathway and risk of osteosarcoma
Article Snippet: .. 2.6 Real‐time PCR Total RNA was isolated from peripheral blood cells using a commercial kit (Tiangen, Beijing, China). ..

Isolation:

Article Title: The Combination of piR-823 and Eukaryotic Initiation Factor 3 B (EIF3B) Activates Hepatic Stellate Cells via Upregulating TGF-β1 in Liver Fibrogenesis
Article Snippet: .. Isolation of RNA and real-time PCR Total RNA was extracted using a Pure Cell/Bacteria kit (Tiangen, Beijing, China). .. The miScript Plant RT Kit (Qiagen, Hilden, Germany) designed for small RNAs 3′end with 2′-O-Me modification was used for small RNA reverse transcription.

Article Title: Inhibition of A20 expression in tumor microenvironment exerts anti-tumor effect through inducing myeloid-derived suppressor cells apoptosis
Article Snippet: .. Real-Time PCR Total RNA was prepared using the RNA Simple Total RNA Kit (TIANGEN), and the isolated RNA was reverse transcribed into cDNA using a PrimeScript™ RT reagent Kit with gDNA Eraser (TaKaRa). .. Primers used were as follows: A20 forward 5′-TTT GCT ACG ACA CTC GGA AC-3′ and reverse 5′-TTC TGA GGA TGT TGC TGA GG-3′; GAPDH forward 5′-CCC AGA AGA CTG TGG ATG G-3′ and reverse 5′-CAC ATT GGG GGT AGG AAC AC-3′.

Article Title: shRNA-Mediated Silencing of Y-Box Binding Protein-1 (YB-1) Suppresses Growth of Neuroblastoma Cell SH-SY5Y In Vitro and In Vivo
Article Snippet: .. RNA isolation and real-time PCR Total RNA of SH-SY5Y parental cells, YB-1-silenced SH-SY5Y cells (SH-shYB-1) and SH-shCON cells was isolated with RNAsimple Total RNA Kit (TIANGEN Biotech, Beijing, China), followed by reverse transcription using Super M-MLV reverse transcriptase (BioTeke, Beijing, China). .. Real-time PCR was performed to determine the levels of YB-1 and internal control β-actin using SYBR Green Master Mix (Solarbio, Beijing, China) and respective primers in an Exicycler 96 quantitative fluorescence analyzer (Bioneer, Daejeon, Korea).

Article Title: SiMYB56 Confers Drought Stress Tolerance in Transgenic Rice by Regulating Lignin Biosynthesis and ABA Signaling Pathway
Article Snippet: .. RNA Isolation and Quantitative Real-Time PCR Total RNA was extracted from seedlings using the Total RNA Extraction Kit (TIANGEN, China). .. The cDNA was synthesized in accordance with Fast Quant RT Super Mix Reverse Transcription Kit instructions (TransGene, Beijing, China).

Article Title: Association between functional polymorphisms in IL‐33/ST2 pathway and risk of osteosarcoma. Association between functional polymorphisms in IL‐33/ST2 pathway and risk of osteosarcoma
Article Snippet: .. 2.6 Real‐time PCR Total RNA was isolated from peripheral blood cells using a commercial kit (Tiangen, Beijing, China). ..

RNA Extraction:

Article Title: Plastid-nucleus communication involves calcium-modulated MAPK signalling
Article Snippet: .. RNA extraction and quantitative real-time PCR Total RNA was extracted from seedlings by RNA extraction kit (Tiangen). .. After DNase I treatment, reverse transcription was conducted using PrimeScript II 1st Strand cDNA Synthesis Kit (Takara) according to the manufacturer's protocol.

Article Title: SiMYB56 Confers Drought Stress Tolerance in Transgenic Rice by Regulating Lignin Biosynthesis and ABA Signaling Pathway
Article Snippet: .. RNA Isolation and Quantitative Real-Time PCR Total RNA was extracted from seedlings using the Total RNA Extraction Kit (TIANGEN, China). .. The cDNA was synthesized in accordance with Fast Quant RT Super Mix Reverse Transcription Kit instructions (TransGene, Beijing, China).

Synthesized:

Article Title: Inhibiting Receptor of Advanced Glycation End Products Attenuates Pressure Overload-Induced Cardiac Dysfunction by Preventing Excessive Autophagy
Article Snippet: .. Quantitative Real-Time PCR Total RNA was extracted from LV tissue by using an RNAprep Pure Tissue Kit (Tiangen, Beijing, China), after which first-strand cDNA was synthesized with a FastKing RT Kit (Tiangen, Beijing, China) and quantitative PCR (qPCR) was performed using SYBR® Premix Ex TaqTM II (Takara, Beijing, China). .. Target gene expression was normalized for that of GAPDH and compared among the groups.

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    tiangen biotech co quantitative real time pcr qrt pcr analysis total rna
    Analysis of gene expressions during four stem swelling stages by <t>qRT-PCR.</t>
    Quantitative Real Time Pcr Qrt Pcr Analysis Total Rna, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 92/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    In vivo gene expression at 12 h (A), 24 h (B), and 36 h (C) relative to the highest level of expression in vitro by real-time <t>PCR</t> analysis . Total bacterial <t>RNA</t> extracted from strain ZY05719 grown in LB broth media was used as the template to assay the in vitro expression levels of the 10 newly identified genes. SPF minipigs were employed as model to study the in vivo expression levels. Pigs were inoculated intravenously with strain ZY05719, and bacterial cells recovered from blood at 12 h, 24 h, and 36 h post-inoculation were considered as in vivo growth bacteria. Total bacterial RNAs extracted from in vivo growth bacterial cells were further analyzed by real-time PCR. To determine whether RNA expression level is induced or upregulated under in vivo conditions, we compared in vivo gene expression with the highest level of expression in vitro . The standard deviations are presented from three pigs each, blood collected at 12, 24 and 48 h. 1, ss-1616 ; 2, trag ; 3, nlpa ; 4, srt ; 5, cwh ; 6, hprk ; 7, ysirk ; 8, ss-1955 ; 9, sdh ; 10, ss-1298 ; gapdh was used as reference gene.
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    tiangen biotech co reverse transcription quantitative real time pcr qrt pcr total rna
    miR-181c-5p inhibited cervical SCC tumor growth in vivo. ( A ) Tumor tissue from subcutaneous xenotransplantation of mice. ( B ) The tumor weight. ( C ) <t>qRT-PCR</t> was used to detect expression levels of miR-181c-5p and GSKIP from the mice. ( D ) IHCs were carried out to detect the impacts of the processes on proliferation, apoptosis, stem cell-like property and EMT from the mice. ( E ) The proportion of Ki67, caspase-3, CD44 or Vimentin positive cells in vivo. The experiments were repeated three times and the data are represented as means ± SD; * P
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    Analysis of gene expressions during four stem swelling stages by qRT-PCR.

    Journal: PeerJ

    Article Title: Identification of the GRAS gene family in the Brassica juncea genome provides insight into its role in stem swelling in stem mustard

    doi: 10.7717/peerj.6682

    Figure Lengend Snippet: Analysis of gene expressions during four stem swelling stages by qRT-PCR.

    Article Snippet: RNA extraction and quantitative real-time PCR (qRT-PCR) analysis Total RNA was extracted from the stem samples at four stem swelling periods (stem diameters were 2 cm, 4 cm, 6 cm, and 8 cm) using the total RNA kit (Tiangen, Beijing, China).

    Techniques: Quantitative RT-PCR

    Expression patterns of PbSSs genes during different development stages, including 15, 39, 63, 22, 87, 101, 125 and 149 (DAF) as determined by qRT-PCR experiment. Mean values and standard deviations (SDs) indicated by error bars. ** significant difference ( p

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: The Sucrose Synthase Gene Family in Chinese Pear (Pyrus bretschneideri Rehd.): Structure, Expression, and Evolution

    doi: 10.3390/molecules23051144

    Figure Lengend Snippet: Expression patterns of PbSSs genes during different development stages, including 15, 39, 63, 22, 87, 101, 125 and 149 (DAF) as determined by qRT-PCR experiment. Mean values and standard deviations (SDs) indicated by error bars. ** significant difference ( p

    Article Snippet: RNA Extraction, cDNA Synthesis and Quantitative Real-Time PCR (qRT-PCR) Total RNA was extracted according to the manufacturer instructions of Tiangen (Beijing, China) plant RNA extraction kit.

    Techniques: Expressing, Quantitative RT-PCR

    In vivo gene expression at 12 h (A), 24 h (B), and 36 h (C) relative to the highest level of expression in vitro by real-time PCR analysis . Total bacterial RNA extracted from strain ZY05719 grown in LB broth media was used as the template to assay the in vitro expression levels of the 10 newly identified genes. SPF minipigs were employed as model to study the in vivo expression levels. Pigs were inoculated intravenously with strain ZY05719, and bacterial cells recovered from blood at 12 h, 24 h, and 36 h post-inoculation were considered as in vivo growth bacteria. Total bacterial RNAs extracted from in vivo growth bacterial cells were further analyzed by real-time PCR. To determine whether RNA expression level is induced or upregulated under in vivo conditions, we compared in vivo gene expression with the highest level of expression in vitro . The standard deviations are presented from three pigs each, blood collected at 12, 24 and 48 h. 1, ss-1616 ; 2, trag ; 3, nlpa ; 4, srt ; 5, cwh ; 6, hprk ; 7, ysirk ; 8, ss-1955 ; 9, sdh ; 10, ss-1298 ; gapdh was used as reference gene.

    Journal: BMC Microbiology

    Article Title: Use of in vivo-induced antigen technology (IVIAT) for the identification of Streptococcus suis serotype 2 in vivo-induced bacterial protein antigens

    doi: 10.1186/1471-2180-9-201

    Figure Lengend Snippet: In vivo gene expression at 12 h (A), 24 h (B), and 36 h (C) relative to the highest level of expression in vitro by real-time PCR analysis . Total bacterial RNA extracted from strain ZY05719 grown in LB broth media was used as the template to assay the in vitro expression levels of the 10 newly identified genes. SPF minipigs were employed as model to study the in vivo expression levels. Pigs were inoculated intravenously with strain ZY05719, and bacterial cells recovered from blood at 12 h, 24 h, and 36 h post-inoculation were considered as in vivo growth bacteria. Total bacterial RNAs extracted from in vivo growth bacterial cells were further analyzed by real-time PCR. To determine whether RNA expression level is induced or upregulated under in vivo conditions, we compared in vivo gene expression with the highest level of expression in vitro . The standard deviations are presented from three pigs each, blood collected at 12, 24 and 48 h. 1, ss-1616 ; 2, trag ; 3, nlpa ; 4, srt ; 5, cwh ; 6, hprk ; 7, ysirk ; 8, ss-1955 ; 9, sdh ; 10, ss-1298 ; gapdh was used as reference gene.

    Article Snippet: Real-time PCR Bacterial total RNA was extracted using RNAprep Bacteria Kit (TIANGEN, China), and residual genomic DNA was removed by using a QIAGEN RNase-Free DNase Set (Qiagen) according to the manufacturer's instructions.

    Techniques: In Vivo, Expressing, In Vitro, Real-time Polymerase Chain Reaction, RNA Expression

    miR-181c-5p inhibited cervical SCC tumor growth in vivo. ( A ) Tumor tissue from subcutaneous xenotransplantation of mice. ( B ) The tumor weight. ( C ) qRT-PCR was used to detect expression levels of miR-181c-5p and GSKIP from the mice. ( D ) IHCs were carried out to detect the impacts of the processes on proliferation, apoptosis, stem cell-like property and EMT from the mice. ( E ) The proportion of Ki67, caspase-3, CD44 or Vimentin positive cells in vivo. The experiments were repeated three times and the data are represented as means ± SD; * P

    Journal: OncoTargets and therapy

    Article Title: MiR-181c-5p Mitigates Tumorigenesis in Cervical Squamous Cell Carcinoma via Targeting Glycogen Synthase Kinase 3β Interaction Protein (GSKIP)

    doi: 10.2147/OTT.S245254

    Figure Lengend Snippet: miR-181c-5p inhibited cervical SCC tumor growth in vivo. ( A ) Tumor tissue from subcutaneous xenotransplantation of mice. ( B ) The tumor weight. ( C ) qRT-PCR was used to detect expression levels of miR-181c-5p and GSKIP from the mice. ( D ) IHCs were carried out to detect the impacts of the processes on proliferation, apoptosis, stem cell-like property and EMT from the mice. ( E ) The proportion of Ki67, caspase-3, CD44 or Vimentin positive cells in vivo. The experiments were repeated three times and the data are represented as means ± SD; * P

    Article Snippet: Reverse Transcription Quantitative Real-Time PCR (qRT-PCR) Total RNA was extracted from cells and tissues with TRNzol Universal Reagent (TIANGEN BIOTECH, Beijing, China) following the manufacturer’s protocol.

    Techniques: In Vivo, Mouse Assay, Quantitative RT-PCR, Expressing