Structured Review

Bio-Rad quantitative real time pcr total rna
DNMT3B modulates exon 4–5–6 skipping by physically interacting with hnRNP-LL and CD45 pre-mRNA. ( A ) Native <t>RNA</t> immunoprecipitation assay (RIP-qPCR) shows DNMT3B interaction with CD45 pre-mRNA in control and ICF1 samples. Binding to HOXC4 mRNA is reported as negative control; ( B ) DNMT3B enrichment at DNA (exons 4, 5 and 6) of CD45 gene by ChIP assay; ( C and D ) Expression level of hnRNP-LL in ICF1 samples and controls by qPCR and western blot, respectively; ( E ) CpG methylation level [log2 (methylated/unmethylated)] at hnRNP-LL promoter in ICF1 samples; ( F ) DNMT3B and hnRNP-LL physically interact in ICF1p2 sample as shown in co-immunoprecipitation experiments; ( G ) Expression level (qPCR) of CD45RABC and the constitutive exon9 using isoform specific oligonucleotides upon siDNMT3B and control siRNAs transfection; ( H ) Semi-quantitative <t>PCR</t> amplification of CD45 splicing isoforms in overexpressing-hnRNP-LL ICF1p2 sample upon siDNMT3B and control siRNAs transfection. * P -adj
Quantitative Real Time Pcr Total Rna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "ICF-specific DNMT3B dysfunction interferes with intragenic regulation of mRNA transcription and alternative splicing"

Article Title: ICF-specific DNMT3B dysfunction interferes with intragenic regulation of mRNA transcription and alternative splicing

Journal: Nucleic Acids Research

doi: 10.1093/nar/gkx163

DNMT3B modulates exon 4–5–6 skipping by physically interacting with hnRNP-LL and CD45 pre-mRNA. ( A ) Native RNA immunoprecipitation assay (RIP-qPCR) shows DNMT3B interaction with CD45 pre-mRNA in control and ICF1 samples. Binding to HOXC4 mRNA is reported as negative control; ( B ) DNMT3B enrichment at DNA (exons 4, 5 and 6) of CD45 gene by ChIP assay; ( C and D ) Expression level of hnRNP-LL in ICF1 samples and controls by qPCR and western blot, respectively; ( E ) CpG methylation level [log2 (methylated/unmethylated)] at hnRNP-LL promoter in ICF1 samples; ( F ) DNMT3B and hnRNP-LL physically interact in ICF1p2 sample as shown in co-immunoprecipitation experiments; ( G ) Expression level (qPCR) of CD45RABC and the constitutive exon9 using isoform specific oligonucleotides upon siDNMT3B and control siRNAs transfection; ( H ) Semi-quantitative PCR amplification of CD45 splicing isoforms in overexpressing-hnRNP-LL ICF1p2 sample upon siDNMT3B and control siRNAs transfection. * P -adj
Figure Legend Snippet: DNMT3B modulates exon 4–5–6 skipping by physically interacting with hnRNP-LL and CD45 pre-mRNA. ( A ) Native RNA immunoprecipitation assay (RIP-qPCR) shows DNMT3B interaction with CD45 pre-mRNA in control and ICF1 samples. Binding to HOXC4 mRNA is reported as negative control; ( B ) DNMT3B enrichment at DNA (exons 4, 5 and 6) of CD45 gene by ChIP assay; ( C and D ) Expression level of hnRNP-LL in ICF1 samples and controls by qPCR and western blot, respectively; ( E ) CpG methylation level [log2 (methylated/unmethylated)] at hnRNP-LL promoter in ICF1 samples; ( F ) DNMT3B and hnRNP-LL physically interact in ICF1p2 sample as shown in co-immunoprecipitation experiments; ( G ) Expression level (qPCR) of CD45RABC and the constitutive exon9 using isoform specific oligonucleotides upon siDNMT3B and control siRNAs transfection; ( H ) Semi-quantitative PCR amplification of CD45 splicing isoforms in overexpressing-hnRNP-LL ICF1p2 sample upon siDNMT3B and control siRNAs transfection. * P -adj

Techniques Used: Immunoprecipitation, Real-time Polymerase Chain Reaction, Binding Assay, Negative Control, Chromatin Immunoprecipitation, Expressing, Western Blot, CpG Methylation Assay, Methylation, Transfection, Amplification

2) Product Images from "NeuroD1 mediates nicotine-induced migration and invasion via regulation of the nicotinic acetylcholine receptor subunits in a subset of neural and neuroendocrine carcinomas"

Article Title: NeuroD1 mediates nicotine-induced migration and invasion via regulation of the nicotinic acetylcholine receptor subunits in a subset of neural and neuroendocrine carcinomas

Journal: Molecular Biology of the Cell

doi: 10.1091/mbc.E13-06-0316

TrkB, a downstream target of NeuroD1, regulates expression of several nAChR subunits in H69 and H82 SCLC and H1155 NSCLC-NE lines. (A) H69, H82, and H1155 cells were stably infected with shTrkB or shControl vectors. Cells were selected by sorting as in Figure 5 . qRT-PCR analysis of TrkB and CHRNA3, A5, A7, and B4, plotted as expression relative to the 18s RNA control. (B) H69 and H82 cells were treated with 10 μM lestaurtinib for 8 h, followed by qRT-PCR analysis of TrkB and CHRNA3, A5, A7, B4, and B2 plotted as described. (C) H82 cells were starved for 8 h in 0.1% FBS. Cells were treated with 1 μM nicotine and 50 ng/ml BDNF for 1 h with or without lestaurtinib. Left, cell lysates immunoblotted with indicated antibodies. Right, blots quantified using LI-COR infrared imaging.
Figure Legend Snippet: TrkB, a downstream target of NeuroD1, regulates expression of several nAChR subunits in H69 and H82 SCLC and H1155 NSCLC-NE lines. (A) H69, H82, and H1155 cells were stably infected with shTrkB or shControl vectors. Cells were selected by sorting as in Figure 5 . qRT-PCR analysis of TrkB and CHRNA3, A5, A7, and B4, plotted as expression relative to the 18s RNA control. (B) H69 and H82 cells were treated with 10 μM lestaurtinib for 8 h, followed by qRT-PCR analysis of TrkB and CHRNA3, A5, A7, B4, and B2 plotted as described. (C) H82 cells were starved for 8 h in 0.1% FBS. Cells were treated with 1 μM nicotine and 50 ng/ml BDNF for 1 h with or without lestaurtinib. Left, cell lysates immunoblotted with indicated antibodies. Right, blots quantified using LI-COR infrared imaging.

Techniques Used: Expressing, Stable Transfection, Infection, Quantitative RT-PCR, Imaging

NeuroD1 regulates the expression of α3 and α5, but not β4, nAChR subunits in SHSY5Y, Clone 5, SCLC, and NSCLC-NE lines. (A) NeuroD1 and α3, α5, and β4 subunits were immunoblotted in 25 μg of lysate protein from SCLC, NSCLC, HBEC3KT, Clone 5, and SHSY5Y cells as in Figure 1A . (B) SHSY5Y was treated with 0.1, 0.5, or 2.5 μM nicotine for 48 h. qRT-PCR of NeuroD1 and CHRNA3, A5, and B4 subunits; 18s RNA control. One of three independent experiments. *** p
Figure Legend Snippet: NeuroD1 regulates the expression of α3 and α5, but not β4, nAChR subunits in SHSY5Y, Clone 5, SCLC, and NSCLC-NE lines. (A) NeuroD1 and α3, α5, and β4 subunits were immunoblotted in 25 μg of lysate protein from SCLC, NSCLC, HBEC3KT, Clone 5, and SHSY5Y cells as in Figure 1A . (B) SHSY5Y was treated with 0.1, 0.5, or 2.5 μM nicotine for 48 h. qRT-PCR of NeuroD1 and CHRNA3, A5, and B4 subunits; 18s RNA control. One of three independent experiments. *** p

Techniques Used: Expressing, Quantitative RT-PCR

Effect of nicotine on expression of NeuroD1 in patient-derived lung cancer, bronchial epithelial, and neuroendocrine cell lines. (A) Patient-derived SCLC, HBEC3KT, Clone 5, SHSY5Y, NTERA2, and rodent PC12 cell lines were lysed, and 25 μg of total protein was resolved on gels and immunoblotted for NeuroD1; GAPDH was the loading control. (B) Immortalized bronchial epithelial cells HBEC3KT, HBEC4KT, and HBEC30KT were treated with 0.25, 0.5, 1, and 2 μM nicotine for 24–48 h. Cells were fixed and immunostained for NeuroD1 and 4′,6-diamidino-2-phenylindole (DAPI). (C) qRT-PCR analysis of NeuroD1 expression in HBEC3KT, Clone5, HBEC30KT, and NSCLC HCC4017 with or without exposure to 0.25 μM nicotine for 48 h, plotted as relative expression. The control was 18s RNA. One of two independent experiments in duplicate. (D) HBEC3KT and Clone 5 cells were treated with 1 μM nicotine for 48 h, and 25 μg of lysate protein was immunoblotted as in A with tubulin as control. Relative expression quantified using LI-COR infrared imaging in three independent experiments; * p
Figure Legend Snippet: Effect of nicotine on expression of NeuroD1 in patient-derived lung cancer, bronchial epithelial, and neuroendocrine cell lines. (A) Patient-derived SCLC, HBEC3KT, Clone 5, SHSY5Y, NTERA2, and rodent PC12 cell lines were lysed, and 25 μg of total protein was resolved on gels and immunoblotted for NeuroD1; GAPDH was the loading control. (B) Immortalized bronchial epithelial cells HBEC3KT, HBEC4KT, and HBEC30KT were treated with 0.25, 0.5, 1, and 2 μM nicotine for 24–48 h. Cells were fixed and immunostained for NeuroD1 and 4′,6-diamidino-2-phenylindole (DAPI). (C) qRT-PCR analysis of NeuroD1 expression in HBEC3KT, Clone5, HBEC30KT, and NSCLC HCC4017 with or without exposure to 0.25 μM nicotine for 48 h, plotted as relative expression. The control was 18s RNA. One of two independent experiments in duplicate. (D) HBEC3KT and Clone 5 cells were treated with 1 μM nicotine for 48 h, and 25 μg of lysate protein was immunoblotted as in A with tubulin as control. Relative expression quantified using LI-COR infrared imaging in three independent experiments; * p

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

Related Articles

Real-time Polymerase Chain Reaction:

Article Title: Mitochondrial Fission Is Required for Angiotensin II-Induced Cardiomyocyte Apoptosis Mediated by a Sirt1-p53 Signaling Pathway
Article Snippet: .. Quantitative real-time PCR Total RNA extracted from cells or heart tissues with TRIZOL was used to perform the reverse transcription with High Capacity cDNA Archive Kit (Bio-Rad, USA). .. Real-time quantitative PCR analysis for Drp1 was performed using TaqMan gene expression assays and the 2−ΔΔCt method with housekeeping gene 18S as the endogenous control.

Article Title: NeuroD1 mediates nicotine-induced migration and invasion via regulation of the nicotinic acetylcholine receptor subunits in a subset of neural and neuroendocrine carcinomas
Article Snippet: .. Quantitative real-time PCR Total RNA from cell lines was isolated with TRI Reagent. cDNA was synthesized using iSCRIPT cDNA Synthesis Kit (Bio-Rad). .. RNA were quantified by real-time PCR (RT-PCR) with iTaq (Bio-Rad) master mix using TaqMan probes for 18s rRNA, NeuroD1, and TrkB (Applied Biosystems) on an ABI 7500 thermocycler.

Article Title: Adora2b Adenosine Receptor Engagement Enhances Regulatory T Cell Abundance during Endotoxin-Induced Pulmonary Inflammation
Article Snippet: .. RNA isolation and Real-time PCR Total RNA was extracted from cells or tissue by Trizol, followed by cDNA synthesis using iScript cDNA Synthesis Kit (Bio-Rad Laboratories, Inc, Munich, Germany) according to the manufacturer's instructions. .. Quantitative real-time reverse transcriptase PCR (qPCR) (iCycler; Bio-Rad Laboratories, Inc.) was performed to measure relative mRNA levels for various transcripts, with qPCR master mix contained 1 µM sense and 1 µM antisense primers with iQ™ SYBR® Green (Bio-Rad Laboratories, Inc.).

Article Title: Notch-Mediated Suppression of TSC2 Expression Regulates Cell Differentiation in the Drosophila Intestinal Stem Cell Lineage
Article Snippet: .. Reverse transcription and real-time PCR Total RNA was extracted using Trizol. cDNAs were synthesized using oligo-dT primers and real-time RTPCR was performed on a BioRad iQ5 detection system (using SYBR Green and ΔΔCt quantification method). .. Gigas and Suppressor of Hairless expression levels were quantified relative to Actin5c expression.

Article Title: Hyperoxia Exacerbates Postnatal Inflammation-Induced Lung Injury in Neonatal BRP-39 Null Mutant Mice Promoting the M1 Macrophage Phenotype
Article Snippet: .. Real-Time PCR Total RNA was reverse-transcribed by the iScript cDNA synthesis kit (Bio-Rad), amplified using SYBR Green PCR Master Mix (Bio-Rad), and detected by the opticon 2 real-time machine (MJ Research). .. ELISA Supernatants and tissue homogenates were analyzed by sandwich ELISA for IL-1β , IL-6, and IL-10, as per manufacturer's (R & D Systems) instructions [ , – ].

Article Title: Different Effects of Human Umbilical Cord Mesenchymal Stem Cells on Glioblastoma Stem Cells by Direct Cell Interaction or Via Released Soluble Factors
Article Snippet: .. Real-Time PCR Total RNA was extracted using the AURUM total RNA Mini Kit (BioRad), according to the manufacturer’s instructions, and reverse transcribed into cDNA using iScript cDNA Synthesis Kit (BioRad). cDNA was amplified using EvaGreen mix (BioRad) on a CF96 Touch real-time PCR (BioRad). ..

Article Title: ICF-specific DNMT3B dysfunction interferes with intragenic regulation of mRNA transcription and alternative splicing
Article Snippet: .. Quantitative real time PCR Total RNA from B-LCLs was reverse-transcribed using iScript cDNA Synthesis kit (Bio-Rad San Diego, CA, USA). .. Quantitative real-time PCR (qPCR) was performed using SsoAdvanced™ universal SYBR® Green supermix (Bio-Rad) on Bio-Rad iCycler according to the manufacturer's protocols.

Amplification:

Article Title: Hyperoxia Exacerbates Postnatal Inflammation-Induced Lung Injury in Neonatal BRP-39 Null Mutant Mice Promoting the M1 Macrophage Phenotype
Article Snippet: .. Real-Time PCR Total RNA was reverse-transcribed by the iScript cDNA synthesis kit (Bio-Rad), amplified using SYBR Green PCR Master Mix (Bio-Rad), and detected by the opticon 2 real-time machine (MJ Research). .. ELISA Supernatants and tissue homogenates were analyzed by sandwich ELISA for IL-1β , IL-6, and IL-10, as per manufacturer's (R & D Systems) instructions [ , – ].

Article Title: Different Effects of Human Umbilical Cord Mesenchymal Stem Cells on Glioblastoma Stem Cells by Direct Cell Interaction or Via Released Soluble Factors
Article Snippet: .. Real-Time PCR Total RNA was extracted using the AURUM total RNA Mini Kit (BioRad), according to the manufacturer’s instructions, and reverse transcribed into cDNA using iScript cDNA Synthesis Kit (BioRad). cDNA was amplified using EvaGreen mix (BioRad) on a CF96 Touch real-time PCR (BioRad). ..

Synthesized:

Article Title: NeuroD1 mediates nicotine-induced migration and invasion via regulation of the nicotinic acetylcholine receptor subunits in a subset of neural and neuroendocrine carcinomas
Article Snippet: .. Quantitative real-time PCR Total RNA from cell lines was isolated with TRI Reagent. cDNA was synthesized using iSCRIPT cDNA Synthesis Kit (Bio-Rad). .. RNA were quantified by real-time PCR (RT-PCR) with iTaq (Bio-Rad) master mix using TaqMan probes for 18s rRNA, NeuroD1, and TrkB (Applied Biosystems) on an ABI 7500 thermocycler.

Article Title: Notch-Mediated Suppression of TSC2 Expression Regulates Cell Differentiation in the Drosophila Intestinal Stem Cell Lineage
Article Snippet: .. Reverse transcription and real-time PCR Total RNA was extracted using Trizol. cDNAs were synthesized using oligo-dT primers and real-time RTPCR was performed on a BioRad iQ5 detection system (using SYBR Green and ΔΔCt quantification method). .. Gigas and Suppressor of Hairless expression levels were quantified relative to Actin5c expression.

Isolation:

Article Title: NeuroD1 mediates nicotine-induced migration and invasion via regulation of the nicotinic acetylcholine receptor subunits in a subset of neural and neuroendocrine carcinomas
Article Snippet: .. Quantitative real-time PCR Total RNA from cell lines was isolated with TRI Reagent. cDNA was synthesized using iSCRIPT cDNA Synthesis Kit (Bio-Rad). .. RNA were quantified by real-time PCR (RT-PCR) with iTaq (Bio-Rad) master mix using TaqMan probes for 18s rRNA, NeuroD1, and TrkB (Applied Biosystems) on an ABI 7500 thermocycler.

Article Title: Adora2b Adenosine Receptor Engagement Enhances Regulatory T Cell Abundance during Endotoxin-Induced Pulmonary Inflammation
Article Snippet: .. RNA isolation and Real-time PCR Total RNA was extracted from cells or tissue by Trizol, followed by cDNA synthesis using iScript cDNA Synthesis Kit (Bio-Rad Laboratories, Inc, Munich, Germany) according to the manufacturer's instructions. .. Quantitative real-time reverse transcriptase PCR (qPCR) (iCycler; Bio-Rad Laboratories, Inc.) was performed to measure relative mRNA levels for various transcripts, with qPCR master mix contained 1 µM sense and 1 µM antisense primers with iQ™ SYBR® Green (Bio-Rad Laboratories, Inc.).

Purification:

Article Title: Multiple roles for a novel RND‐type efflux system in Acinetobacter baumannii AB5075Multiple roles for a novel RND‐type efflux system in Acinetobacter baumannii AB5075
Article Snippet: .. 2.11 Quantitative real‐time PCR Total RNA (1 μg) purified from strains AB5075 with or without plasmid and ΔarpR were converted into cDNA by using the iScript cDNA synthesis kit (Bio‐Rad) with random primers and SuperScript III reverse transcriptase (Invitrogen). .. Cycling parameters for cDNA synthesis were as follows: 25°C for 5 min, 42°C for 45 min, and 85°C for 5 min. cDNA reactions were then diluted 1:10 with sterile H2 O supplemented with 10 μg/ml yeast t‐RNA (Roche).

SYBR Green Assay:

Article Title: Notch-Mediated Suppression of TSC2 Expression Regulates Cell Differentiation in the Drosophila Intestinal Stem Cell Lineage
Article Snippet: .. Reverse transcription and real-time PCR Total RNA was extracted using Trizol. cDNAs were synthesized using oligo-dT primers and real-time RTPCR was performed on a BioRad iQ5 detection system (using SYBR Green and ΔΔCt quantification method). .. Gigas and Suppressor of Hairless expression levels were quantified relative to Actin5c expression.

Article Title: Hyperoxia Exacerbates Postnatal Inflammation-Induced Lung Injury in Neonatal BRP-39 Null Mutant Mice Promoting the M1 Macrophage Phenotype
Article Snippet: .. Real-Time PCR Total RNA was reverse-transcribed by the iScript cDNA synthesis kit (Bio-Rad), amplified using SYBR Green PCR Master Mix (Bio-Rad), and detected by the opticon 2 real-time machine (MJ Research). .. ELISA Supernatants and tissue homogenates were analyzed by sandwich ELISA for IL-1β , IL-6, and IL-10, as per manufacturer's (R & D Systems) instructions [ , – ].

Reverse Transcription Polymerase Chain Reaction:

Article Title: Notch-Mediated Suppression of TSC2 Expression Regulates Cell Differentiation in the Drosophila Intestinal Stem Cell Lineage
Article Snippet: .. Reverse transcription and real-time PCR Total RNA was extracted using Trizol. cDNAs were synthesized using oligo-dT primers and real-time RTPCR was performed on a BioRad iQ5 detection system (using SYBR Green and ΔΔCt quantification method). .. Gigas and Suppressor of Hairless expression levels were quantified relative to Actin5c expression.

Polymerase Chain Reaction:

Article Title: Hyperoxia Exacerbates Postnatal Inflammation-Induced Lung Injury in Neonatal BRP-39 Null Mutant Mice Promoting the M1 Macrophage Phenotype
Article Snippet: .. Real-Time PCR Total RNA was reverse-transcribed by the iScript cDNA synthesis kit (Bio-Rad), amplified using SYBR Green PCR Master Mix (Bio-Rad), and detected by the opticon 2 real-time machine (MJ Research). .. ELISA Supernatants and tissue homogenates were analyzed by sandwich ELISA for IL-1β , IL-6, and IL-10, as per manufacturer's (R & D Systems) instructions [ , – ].

Plasmid Preparation:

Article Title: Multiple roles for a novel RND‐type efflux system in Acinetobacter baumannii AB5075Multiple roles for a novel RND‐type efflux system in Acinetobacter baumannii AB5075
Article Snippet: .. 2.11 Quantitative real‐time PCR Total RNA (1 μg) purified from strains AB5075 with or without plasmid and ΔarpR were converted into cDNA by using the iScript cDNA synthesis kit (Bio‐Rad) with random primers and SuperScript III reverse transcriptase (Invitrogen). .. Cycling parameters for cDNA synthesis were as follows: 25°C for 5 min, 42°C for 45 min, and 85°C for 5 min. cDNA reactions were then diluted 1:10 with sterile H2 O supplemented with 10 μg/ml yeast t‐RNA (Roche).

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    Bio-Rad quantitative real time reverse transcription rt pcr purified total rna
    Pearson correlation value between the log2-transformed fold changes of <t>RNA-Seq</t> data and <t>qRT-PCR</t> validation. Each dot (in blue) represents a data point for the selected genes and each corresponding sample (4 genes and 8 replicates in control (D0) and later time point, D1 and D14, respectively). And the dashed lines are 95% confidence interval.
    Quantitative Real Time Reverse Transcription Rt Pcr Purified Total Rna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time reverse transcription rt pcr purified total rna/product/Bio-Rad
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    quantitative real time reverse transcription rt pcr purified total rna - by Bioz Stars, 2020-08
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    93
    Bio-Rad real time pcr analysis total rna
    High temperature induces endoplasmic reticulum (ER) stress-mediated cell death. ( a ) Protein expression under condition of hyperthermia was measured. Cell lysates were collected at indicated times following incubation at 42 °C or tunicamycin (Tm) (1 μg/mL) treatment for western blot analysis. ( b ) Effect of heat stress on the unfolded protein response (UPR) gene expression. Quantitative <t>RT-PCR</t> was performed using total <t>RNA</t> extracted from mouse embryonic fibroblasts (MEFs) which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( c ) KDEL and DAPI immunofluorescence staining. KDEL and DAPI staining were performed using MEFs incubated at 42 °C or treated with thapsigargin (Tg) (300 nM) for 12 h. Representative images are shown. Scale bar represents 100 μm. ( d ) Cell viability was measured in MEFs treated with Tg (300 nM), Tm (1 μg/mL), or incubated at 42 °C for the indicated time periods. ( e ) Cell viability was measured in MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of tauroursodeoxycholic acid (TUDCA) (2.4 mM). ** p
    Real Time Pcr Analysis Total Rna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time pcr analysis total rna/product/Bio-Rad
    Average 93 stars, based on 15 article reviews
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    Bio-Rad total rna
    Immune response of hVECs analyzed by <t>qRT-PCR</t> after incubation with siICAM-1, scrRNA or Lipofectamine ® 2000 alone, PLGA 1–3 coatings and PLGA 1-3 alone. After 24 h incubation of coated slides with hVECs, <t>RNA</t> was isolated for qRT-PCR. The relative expression of ( a ) CXCL-7, ( b ) CXCL-10, ( c ) OAS, and ( d ) STAT-1 was compared to untreated cells. Another assay was the transfection of hVECs with dsRNA (2 and 5 µg): ( a’ ) relative expression of CXCL-7, ( b’ ) relative expression of CXCL-10, ( c’ ) relative expression of OAS, and ( d’ ) relative expression of STAT-1. The expression of untreated cells was set to one. Each bar represents the mean ± standard error (SEM) of n = 4. The dsRNA was tested only once, n = 1. p = PLGA.
    Total Rna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 2087 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/total rna/product/Bio-Rad
    Average 94 stars, based on 2087 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    Pearson correlation value between the log2-transformed fold changes of RNA-Seq data and qRT-PCR validation. Each dot (in blue) represents a data point for the selected genes and each corresponding sample (4 genes and 8 replicates in control (D0) and later time point, D1 and D14, respectively). And the dashed lines are 95% confidence interval.

    Journal: Scientific Reports

    Article Title: Temporal dynamics in meta longitudinal RNA-Seq data

    doi: 10.1038/s41598-018-37397-7

    Figure Lengend Snippet: Pearson correlation value between the log2-transformed fold changes of RNA-Seq data and qRT-PCR validation. Each dot (in blue) represents a data point for the selected genes and each corresponding sample (4 genes and 8 replicates in control (D0) and later time point, D1 and D14, respectively). And the dashed lines are 95% confidence interval.

    Article Snippet: Quantitative Real-time Reverse Transcription (RT) PCR Purified total RNA samples were converted to cDNA using an iScript Advanced cDNA Synthesis Kit (Bio-Rad, Hercules, CA, USA).

    Techniques: Transformation Assay, RNA Sequencing Assay, Quantitative RT-PCR

    High temperature induces endoplasmic reticulum (ER) stress-mediated cell death. ( a ) Protein expression under condition of hyperthermia was measured. Cell lysates were collected at indicated times following incubation at 42 °C or tunicamycin (Tm) (1 μg/mL) treatment for western blot analysis. ( b ) Effect of heat stress on the unfolded protein response (UPR) gene expression. Quantitative RT-PCR was performed using total RNA extracted from mouse embryonic fibroblasts (MEFs) which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( c ) KDEL and DAPI immunofluorescence staining. KDEL and DAPI staining were performed using MEFs incubated at 42 °C or treated with thapsigargin (Tg) (300 nM) for 12 h. Representative images are shown. Scale bar represents 100 μm. ( d ) Cell viability was measured in MEFs treated with Tg (300 nM), Tm (1 μg/mL), or incubated at 42 °C for the indicated time periods. ( e ) Cell viability was measured in MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of tauroursodeoxycholic acid (TUDCA) (2.4 mM). ** p

    Journal: Cells

    Article Title: Modulation of Protein Synthesis by eIF2α Phosphorylation Protects Cell from Heat Stress-Mediated Apoptosis

    doi: 10.3390/cells7120254

    Figure Lengend Snippet: High temperature induces endoplasmic reticulum (ER) stress-mediated cell death. ( a ) Protein expression under condition of hyperthermia was measured. Cell lysates were collected at indicated times following incubation at 42 °C or tunicamycin (Tm) (1 μg/mL) treatment for western blot analysis. ( b ) Effect of heat stress on the unfolded protein response (UPR) gene expression. Quantitative RT-PCR was performed using total RNA extracted from mouse embryonic fibroblasts (MEFs) which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( c ) KDEL and DAPI immunofluorescence staining. KDEL and DAPI staining were performed using MEFs incubated at 42 °C or treated with thapsigargin (Tg) (300 nM) for 12 h. Representative images are shown. Scale bar represents 100 μm. ( d ) Cell viability was measured in MEFs treated with Tg (300 nM), Tm (1 μg/mL), or incubated at 42 °C for the indicated time periods. ( e ) Cell viability was measured in MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of tauroursodeoxycholic acid (TUDCA) (2.4 mM). ** p

    Article Snippet: RNA Extraction and Real-Time PCR Analysis Total RNA was extracted from cells using Trizol (Gene All, Ribo-Ex, Seoul, Korea) and cDNA was synthesized using an iScript cDNA synthesis kit (Bio-Rad, Hercules, CA, USA, BR170-8891).

    Techniques: Expressing, Incubation, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining

    The ATF6α pathway does not protect cells from heat stress-mediated death. ( a ) Quantitative RT-PCR was performed using total RNA extracted from Atf6α −/− and Atf6α +/+ MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( b ) Protein expression under conditions of hyperthermia was measured. Following 42 °C incubation, cell lysates were collected at indicated times for western blot analysis. ( c ) Cell viability was measured in Atf6α −/− and Atf6α +/+ MEFs following incubation at 42 °C for indicated time periods. ( d ) Caspase 3/7 activity was detected in Atf6α −/− and Atf6α +/+ MEFs which were incubated at 42 °C for 12 h. ** p

    Journal: Cells

    Article Title: Modulation of Protein Synthesis by eIF2α Phosphorylation Protects Cell from Heat Stress-Mediated Apoptosis

    doi: 10.3390/cells7120254

    Figure Lengend Snippet: The ATF6α pathway does not protect cells from heat stress-mediated death. ( a ) Quantitative RT-PCR was performed using total RNA extracted from Atf6α −/− and Atf6α +/+ MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( b ) Protein expression under conditions of hyperthermia was measured. Following 42 °C incubation, cell lysates were collected at indicated times for western blot analysis. ( c ) Cell viability was measured in Atf6α −/− and Atf6α +/+ MEFs following incubation at 42 °C for indicated time periods. ( d ) Caspase 3/7 activity was detected in Atf6α −/− and Atf6α +/+ MEFs which were incubated at 42 °C for 12 h. ** p

    Article Snippet: RNA Extraction and Real-Time PCR Analysis Total RNA was extracted from cells using Trizol (Gene All, Ribo-Ex, Seoul, Korea) and cDNA was synthesized using an iScript cDNA synthesis kit (Bio-Rad, Hercules, CA, USA, BR170-8891).

    Techniques: Quantitative RT-PCR, Incubation, Expressing, Western Blot, Activity Assay

    eIF2α phosphorylation is required to protect cells from heat stress-mediated death. ( a ) Protein expression was measured under conditions of hyperthermia. Cell lysates were collected from Eif2α S/S and Eif2α A/A MEFs at indicated times for western blot analysis following incubation at 42 °C. ( b ) Quantitative RT-PCR was performed using total RNA extracted from Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( c ) Cell viability was measured in Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for indicated times. ( d ) Caspase 3/7 activity was detected in Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for 12 h. ( e ) Cell viability was measured in MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of guanabenz (GA, 1 μM) or ISRIB (5 μM). ( f ) Cell viability was measured in Perk +/+ and Perk −/− MEFs which were incubated at 42 °C for 12 and 24 h. ( g ) Cell viability was measured in MEFs which were incubated at 42 °C for 12 h in the presence or absence of GSK2606414 (100 nM). * p

    Journal: Cells

    Article Title: Modulation of Protein Synthesis by eIF2α Phosphorylation Protects Cell from Heat Stress-Mediated Apoptosis

    doi: 10.3390/cells7120254

    Figure Lengend Snippet: eIF2α phosphorylation is required to protect cells from heat stress-mediated death. ( a ) Protein expression was measured under conditions of hyperthermia. Cell lysates were collected from Eif2α S/S and Eif2α A/A MEFs at indicated times for western blot analysis following incubation at 42 °C. ( b ) Quantitative RT-PCR was performed using total RNA extracted from Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM (n = 3 independent experiments). ( c ) Cell viability was measured in Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for indicated times. ( d ) Caspase 3/7 activity was detected in Eif2α S/S and Eif2α A/A MEFs which were incubated at 42 °C for 12 h. ( e ) Cell viability was measured in MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of guanabenz (GA, 1 μM) or ISRIB (5 μM). ( f ) Cell viability was measured in Perk +/+ and Perk −/− MEFs which were incubated at 42 °C for 12 and 24 h. ( g ) Cell viability was measured in MEFs which were incubated at 42 °C for 12 h in the presence or absence of GSK2606414 (100 nM). * p

    Article Snippet: RNA Extraction and Real-Time PCR Analysis Total RNA was extracted from cells using Trizol (Gene All, Ribo-Ex, Seoul, Korea) and cDNA was synthesized using an iScript cDNA synthesis kit (Bio-Rad, Hercules, CA, USA, BR170-8891).

    Techniques: Expressing, Western Blot, Incubation, Quantitative RT-PCR, Activity Assay

    The IRE1α pathway does not protect cells from heat stress-mediated death. ( a ) Quantitative RT-PCR was performed using total RNA, extracted from Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM. (n = 3 independent experiments). ( b ) Protein expression under conditions of hyperthermia was measured. Following incubation at 42 °C, cell lysates were collected at the indicated times for western blot analysis. ( c ) Cell viability was measured in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods. ( d ) Caspase 3/7 activity was detected in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for 12 h. ( e ) Expression of sXBP1 (spliced form) and tXBP (total form) were measured using MEFs treated with 4μ8c at indicated doses in the presence or absence of Tg (300 nM) for 12 h. ( f ) Protein expression under conditions of hyperthermia was measured in the presence or absence of 4μ8c (20 μM). ( g ) Cell viability was measured in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of 4μ8c (20 μM). * p

    Journal: Cells

    Article Title: Modulation of Protein Synthesis by eIF2α Phosphorylation Protects Cell from Heat Stress-Mediated Apoptosis

    doi: 10.3390/cells7120254

    Figure Lengend Snippet: The IRE1α pathway does not protect cells from heat stress-mediated death. ( a ) Quantitative RT-PCR was performed using total RNA, extracted from Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods. Data are presented as means ± SEM. (n = 3 independent experiments). ( b ) Protein expression under conditions of hyperthermia was measured. Following incubation at 42 °C, cell lysates were collected at the indicated times for western blot analysis. ( c ) Cell viability was measured in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods. ( d ) Caspase 3/7 activity was detected in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for 12 h. ( e ) Expression of sXBP1 (spliced form) and tXBP (total form) were measured using MEFs treated with 4μ8c at indicated doses in the presence or absence of Tg (300 nM) for 12 h. ( f ) Protein expression under conditions of hyperthermia was measured in the presence or absence of 4μ8c (20 μM). ( g ) Cell viability was measured in Ire1α −/− and Ire1α +/+ MEFs which were incubated at 42 °C for indicated time periods in the presence or absence of 4μ8c (20 μM). * p

    Article Snippet: RNA Extraction and Real-Time PCR Analysis Total RNA was extracted from cells using Trizol (Gene All, Ribo-Ex, Seoul, Korea) and cDNA was synthesized using an iScript cDNA synthesis kit (Bio-Rad, Hercules, CA, USA, BR170-8891).

    Techniques: Quantitative RT-PCR, Incubation, Expressing, Western Blot, Activity Assay

    UGT mRNA expression in primary human melanocytes and melanoma. (A) RT-PCR analysis of total RNA from human melanocytes for UGT family members. Primer sets for the UGT2Bs were designed to be specific to each isoform while a single primer set directed against the common region of all UGT1As was used to detect UGT1A expression. Total RNA from human liver was used as a control with UGT2B7 primers. Arrows indicate DNA bands of expected size whose sequence was confirmed. (B) Control RT-PCR analysis of total RNA from liver using indicated primers sets. (C) RT-PCR analysis of total RNA from the human melanoma cell line WM115 using indicated primers sets. Arrows indicate bands that were excised and sequenced. Band in UGT2B4 lane was found to be UGT2B7 by sequencing while UGT2B10 and UGT2B15 bands were confirmed to be expected UGTs. (D) RT-PCR analysis of total RNA from the human melanoma cell line WM3211. GAPDH was used as a positive control here to ensure cDNA quality. (E) Real-time PCR using TaqMan assays against indicated UGTs. ND = not detected.

    Journal: PLoS ONE

    Article Title: Anti-Cancer Drugs Elicit Re-Expression of UDP-Glucuronosyltransferases in Melanoma Cells

    doi: 10.1371/journal.pone.0047696

    Figure Lengend Snippet: UGT mRNA expression in primary human melanocytes and melanoma. (A) RT-PCR analysis of total RNA from human melanocytes for UGT family members. Primer sets for the UGT2Bs were designed to be specific to each isoform while a single primer set directed against the common region of all UGT1As was used to detect UGT1A expression. Total RNA from human liver was used as a control with UGT2B7 primers. Arrows indicate DNA bands of expected size whose sequence was confirmed. (B) Control RT-PCR analysis of total RNA from liver using indicated primers sets. (C) RT-PCR analysis of total RNA from the human melanoma cell line WM115 using indicated primers sets. Arrows indicate bands that were excised and sequenced. Band in UGT2B4 lane was found to be UGT2B7 by sequencing while UGT2B10 and UGT2B15 bands were confirmed to be expected UGTs. (D) RT-PCR analysis of total RNA from the human melanoma cell line WM3211. GAPDH was used as a positive control here to ensure cDNA quality. (E) Real-time PCR using TaqMan assays against indicated UGTs. ND = not detected.

    Article Snippet: Total RNA Isolation, Reverse Transcription and PCR Total RNA was isolated from cells using the Arum total RNA mini Kit (BioRad) according to companies provided protocol.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Sequencing, Positive Control, Real-time Polymerase Chain Reaction

    Immune response of hVECs analyzed by qRT-PCR after incubation with siICAM-1, scrRNA or Lipofectamine ® 2000 alone, PLGA 1–3 coatings and PLGA 1-3 alone. After 24 h incubation of coated slides with hVECs, RNA was isolated for qRT-PCR. The relative expression of ( a ) CXCL-7, ( b ) CXCL-10, ( c ) OAS, and ( d ) STAT-1 was compared to untreated cells. Another assay was the transfection of hVECs with dsRNA (2 and 5 µg): ( a’ ) relative expression of CXCL-7, ( b’ ) relative expression of CXCL-10, ( c’ ) relative expression of OAS, and ( d’ ) relative expression of STAT-1. The expression of untreated cells was set to one. Each bar represents the mean ± standard error (SEM) of n = 4. The dsRNA was tested only once, n = 1. p = PLGA.

    Journal: Pharmaceuticals

    Article Title: RNA-Eluting Surfaces for the Modulation of Gene Expression as A Novel Stent Concept

    doi: 10.3390/ph10010023

    Figure Lengend Snippet: Immune response of hVECs analyzed by qRT-PCR after incubation with siICAM-1, scrRNA or Lipofectamine ® 2000 alone, PLGA 1–3 coatings and PLGA 1-3 alone. After 24 h incubation of coated slides with hVECs, RNA was isolated for qRT-PCR. The relative expression of ( a ) CXCL-7, ( b ) CXCL-10, ( c ) OAS, and ( d ) STAT-1 was compared to untreated cells. Another assay was the transfection of hVECs with dsRNA (2 and 5 µg): ( a’ ) relative expression of CXCL-7, ( b’ ) relative expression of CXCL-10, ( c’ ) relative expression of OAS, and ( d’ ) relative expression of STAT-1. The expression of untreated cells was set to one. Each bar represents the mean ± standard error (SEM) of n = 4. The dsRNA was tested only once, n = 1. p = PLGA.

    Article Snippet: Quantitative Real-Time PCR (qRT-PCR) First of all, total RNA from PLGA/RNA transfected EA.hy926 and hVECs was isolated using Aurum™ total RNA mini kit (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

    Techniques: Quantitative RT-PCR, Incubation, Isolation, Expressing, Transfection

    Sequence of 5′-RLM-RACE-PCR product (Inner PCR with Inner Primer and RACE1) of PLGA-ICAM-1 siRNA–treated EA.hy926 cells. The sequence shows the siRNA cleavage site of the ICAM-1 mRNA at bp 1818 (126–144) and the RNA Adapter sequence (145–161).

    Journal: Pharmaceuticals

    Article Title: RNA-Eluting Surfaces for the Modulation of Gene Expression as A Novel Stent Concept

    doi: 10.3390/ph10010023

    Figure Lengend Snippet: Sequence of 5′-RLM-RACE-PCR product (Inner PCR with Inner Primer and RACE1) of PLGA-ICAM-1 siRNA–treated EA.hy926 cells. The sequence shows the siRNA cleavage site of the ICAM-1 mRNA at bp 1818 (126–144) and the RNA Adapter sequence (145–161).

    Article Snippet: Quantitative Real-Time PCR (qRT-PCR) First of all, total RNA from PLGA/RNA transfected EA.hy926 and hVECs was isolated using Aurum™ total RNA mini kit (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

    Techniques: Sequencing, Polymerase Chain Reaction

    Illustration of the RNA Interference mechanism and the 5´-RLM-RACE PCR technique. ( a ) Shows the predicted siRNA cleavage site in the ICAM-1 mRNA. ( b ) Illustrates the RNAi mechanism in the cytoplasm of EA.hy926 cells. ( c ) Demonstrates the 5´-RLM RACE PCR technique, starting with total RNA Isolation, then the RNA adapter is ligated to cleaved mRNA strands. Afterwards RNA is reverse transcribed gene-specifically, with specific first-strand synthesis primers for ICAM-1. Subsequently, Outer PCR and Inner PCR with gene-specific primers (RACE 1, 2, and 3) and adapter-specific primers (Outer and Inner Primer) are conducted. Only if siRNA-mediated cleavage occurs specific products are produced with primer combinations of Inner Primer and RACE 1. These PCR products are then sequenced to prove siRNA-specific degradation.

    Journal: Pharmaceuticals

    Article Title: RNA-Eluting Surfaces for the Modulation of Gene Expression as A Novel Stent Concept

    doi: 10.3390/ph10010023

    Figure Lengend Snippet: Illustration of the RNA Interference mechanism and the 5´-RLM-RACE PCR technique. ( a ) Shows the predicted siRNA cleavage site in the ICAM-1 mRNA. ( b ) Illustrates the RNAi mechanism in the cytoplasm of EA.hy926 cells. ( c ) Demonstrates the 5´-RLM RACE PCR technique, starting with total RNA Isolation, then the RNA adapter is ligated to cleaved mRNA strands. Afterwards RNA is reverse transcribed gene-specifically, with specific first-strand synthesis primers for ICAM-1. Subsequently, Outer PCR and Inner PCR with gene-specific primers (RACE 1, 2, and 3) and adapter-specific primers (Outer and Inner Primer) are conducted. Only if siRNA-mediated cleavage occurs specific products are produced with primer combinations of Inner Primer and RACE 1. These PCR products are then sequenced to prove siRNA-specific degradation.

    Article Snippet: Quantitative Real-Time PCR (qRT-PCR) First of all, total RNA from PLGA/RNA transfected EA.hy926 and hVECs was isolated using Aurum™ total RNA mini kit (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

    Techniques: Polymerase Chain Reaction, Isolation, Produced