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Bio-Rad superscript iii reverse transcriptase
PPARs’ <t>mRNA</t> and protein expression during maturation and embryo development. ( A ) RT-PCR-based PPARα, PPARγ, and PPARδ expression at the COCs (cumulus oocyte complexes) (10 per sample), MII oocyte (20 per sample), two-cell embryo (20 per sample), 3.5-day embryo (10 per sample) and day-8 blastocyst stages (five per sample). ( B ) Relative mRNA expressions of PPARα, PPARγ and PPARδ from the COCs stage to day-8 blastocysts. ( C ) Development stage-dependent immunofluorescent expression of PPARα and PPARγ ( n = 15 per sample). ( D ) Immunofluorescent co-localization of PPARδ and PEPCK from COCs to day-8 blastocysts ( n = 15 per sample). The experiments were repeated <t>three</t> times, and the data are shown here as mean ± S.E.M. N.S., not significant. Significance: * p
Superscript Iii Reverse Transcriptase, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "The PPARδ Agonist GW501516 Improves Lipolytic/Lipogenic Balance through CPT1 and PEPCK during the Development of Pre-Implantation Bovine Embryos"

Article Title: The PPARδ Agonist GW501516 Improves Lipolytic/Lipogenic Balance through CPT1 and PEPCK during the Development of Pre-Implantation Bovine Embryos

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms20236066

PPARs’ mRNA and protein expression during maturation and embryo development. ( A ) RT-PCR-based PPARα, PPARγ, and PPARδ expression at the COCs (cumulus oocyte complexes) (10 per sample), MII oocyte (20 per sample), two-cell embryo (20 per sample), 3.5-day embryo (10 per sample) and day-8 blastocyst stages (five per sample). ( B ) Relative mRNA expressions of PPARα, PPARγ and PPARδ from the COCs stage to day-8 blastocysts. ( C ) Development stage-dependent immunofluorescent expression of PPARα and PPARγ ( n = 15 per sample). ( D ) Immunofluorescent co-localization of PPARδ and PEPCK from COCs to day-8 blastocysts ( n = 15 per sample). The experiments were repeated three times, and the data are shown here as mean ± S.E.M. N.S., not significant. Significance: * p
Figure Legend Snippet: PPARs’ mRNA and protein expression during maturation and embryo development. ( A ) RT-PCR-based PPARα, PPARγ, and PPARδ expression at the COCs (cumulus oocyte complexes) (10 per sample), MII oocyte (20 per sample), two-cell embryo (20 per sample), 3.5-day embryo (10 per sample) and day-8 blastocyst stages (five per sample). ( B ) Relative mRNA expressions of PPARα, PPARγ and PPARδ from the COCs stage to day-8 blastocysts. ( C ) Development stage-dependent immunofluorescent expression of PPARα and PPARγ ( n = 15 per sample). ( D ) Immunofluorescent co-localization of PPARδ and PEPCK from COCs to day-8 blastocysts ( n = 15 per sample). The experiments were repeated three times, and the data are shown here as mean ± S.E.M. N.S., not significant. Significance: * p

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

PPARδ reversibility and lipid metabolism in bovine embryos. ( A ) Immunofluorescent co-localization of PEPCK and CPT1 in control-, GSK3787-, and GW501516 IVC-treated day-8 blastocysts (15 per group). The original magnification is ×200. ( B ) RT-qPCR-based mRNA quantification of ATGL, LMF1, LMF2, and LPL in day-8 blastocysts (five per sample). ( C ) Western blot analysis of SIRT1 and p-mTOR in control-, GSK3787-, and GW501516 IVC-treated blastocysts (20 per group). The experiments were repeated three times, and the data are shown here as mean ± S.E.M. * p
Figure Legend Snippet: PPARδ reversibility and lipid metabolism in bovine embryos. ( A ) Immunofluorescent co-localization of PEPCK and CPT1 in control-, GSK3787-, and GW501516 IVC-treated day-8 blastocysts (15 per group). The original magnification is ×200. ( B ) RT-qPCR-based mRNA quantification of ATGL, LMF1, LMF2, and LPL in day-8 blastocysts (five per sample). ( C ) Western blot analysis of SIRT1 and p-mTOR in control-, GSK3787-, and GW501516 IVC-treated blastocysts (20 per group). The experiments were repeated three times, and the data are shown here as mean ± S.E.M. * p

Techniques Used: Quantitative RT-PCR, Western Blot

Related Articles

Real-time Polymerase Chain Reaction:

Article Title: RAS P21 Protein Activator 3 (RASA3) specifically promotes pathogenic T helper 17 cell generation by repressing T helper 2-biased programs
Article Snippet: For qRT-PCR analysis, total RNA was extracted from lymphocytes using TRizol reagent (Invitrogen) and reverse-transcribed into cDNA with Superscript III reverse transcriptase (Bio-Rad) per manufacturer’s protocols. .. Quantitative PCR (qPCR) was performed on QuantStudio® 6 Flex Real-Time PCR Syst em (ThermoFisher Scientific).

Article Title: Collaborative interactions between type-2 innate lymphoid cells and Ag-specific CD4+Th2 cells exacerbate murine allergic airway diseases with prominent eosinophilia
Article Snippet: Paragraph title: RNA isolation and quantitative real-time PCR analysis ... RNA from sorted cell populations was isolated with an RNeasy Plus Mini kit (Qiagen), and cDNA templates were synthesized with SuperScript III reverse transcriptase (BioRad).

Article Title: Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W]Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W] [OA]
Article Snippet: .. For P. abies , total RNA was isolated from needles or buds following the protocol described by with minor modifications, while for Arabidopsis, the Qiagen RNeasy Mini kit was used in accordance with the manufacturer’s recommendations. cDNAs were synthesized from 0.5 μg of total RNA using SuperScript III reverse transcriptase and random hexamer primers. qRT-PCR was performed according to or on a MyiQ Real-Time PCR Detection system (Bio-Rad) with the thermal conditions 95°C for 7 min followed by 40 cycles of 95°C for 10 s and 60°C for 30 s. Each reaction was performed in duplicate containing 12 μL of DyNAmo Flash SYBRGreen (DyNAmo Flash SYBRGreen qPCR kit; Finnzymes), 0.5 m m of each primer, and 4.75 μL of cDNA (diluted 1:100) in a total volume of 23.75 μL. ..

Article Title: Endotrophin triggers adipose tissue fibrosis and metabolic dysfunction
Article Snippet: .. The purity and quantity of the mRNAs were determined by absorbance at 260/280 nm. cDNAs were prepared by reverse transcribing 500 ng of total mRNAs with a kit containing Superscript III reverse transcriptase and oligo (dT)20 (Bio-Rad). qPCR reactions were carried out on an ABI Prism 7900 HT sequence detection system (Applied Biosystems). ..

Article Title: BPTF is essential for T cell homeostasis and function
Article Snippet: Total RNA was extracted from T cells with TRIzol reagent according to the manufacturer’s instructions (Invitrogen) and was reverse-transcribed into c-DNA with Superscript III reverse transcriptase (Bio-Rad). .. Quantitative PCR was performed on QuantStudio® 6 Flex Real-Time PCR System.

Article Title: Kr?ppel-like factor 4 is induced by rapamycin and mediates the anti-proliferative effect of rapamycin in rat carotid arteries after balloon injury
Article Snippet: .. Total RNA was isolated from VSMCs and arterial segments with Trizol Reagent (Invitrogen, Grand Island, NY, USA). cDNA was synthesized with the use of Superscript III reverse transcriptase and oligo-(dT) primer. qPCR was performed with the iQ™ SYBR Green PCR Supermix in the DNA Engine Opticon real-time system (Bio-Rad, Hercules, CA, USA), with GAPDH used as an internal control. ..

Article Title: RAS P21 Protein Activator 3 (RASA3) specifically promotes pathogenic T helper 17 cell generation by repressing T helper 2-biased programs
Article Snippet: For qRT-PCR analysis, total RNA was extracted from lymphocytes using TRizol reagent (Invitrogen) and reverse-transcribed into cDNA with Superscript III reverse transcriptase (Bio-Rad) per manufacturer’s protocols. .. Quantitative PCR (qPCR) was performed on QuantStudio® 6 Flex Real-Time PCR Syst em (ThermoFisher Scientific).

RNA Extraction:

Article Title: Endotrophin triggers adipose tissue fibrosis and metabolic dysfunction
Article Snippet: Total RNAs were extracted in Trizol (Invitrogen, Carlsbad, CA) using a TissueLyser (Qiagen, Valenica, CA) and then isolated with the RNeasy RNA extraction kit (Qiagen) following the company's protocol. .. The purity and quantity of the mRNAs were determined by absorbance at 260/280 nm. cDNAs were prepared by reverse transcribing 500 ng of total mRNAs with a kit containing Superscript III reverse transcriptase and oligo (dT)20 (Bio-Rad). qPCR reactions were carried out on an ABI Prism 7900 HT sequence detection system (Applied Biosystems).

Sequencing:

Article Title: Endotrophin triggers adipose tissue fibrosis and metabolic dysfunction
Article Snippet: .. The purity and quantity of the mRNAs were determined by absorbance at 260/280 nm. cDNAs were prepared by reverse transcribing 500 ng of total mRNAs with a kit containing Superscript III reverse transcriptase and oligo (dT)20 (Bio-Rad). qPCR reactions were carried out on an ABI Prism 7900 HT sequence detection system (Applied Biosystems). ..

Article Title: Mov10 suppresses retroelements and regulates neuronal development and function in the developing brain
Article Snippet: The RNA was extracted with phenol-chloroform and precipitated in ethanol and converted into cDNA using Oligo dT primer and SuperScript III Reverse Transcriptase. qRT-PCR was performed with iQ SYBR Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA) using a StepOnePlus RT PCR machine (Applied Biosystems) with gene-specific primers. .. For the brain IP-RNA sequencing, WT brains were homogenized in the same manner as described above but were not UV cross-linked.

Sample Prep:

Article Title: RAS P21 Protein Activator 3 (RASA3) specifically promotes pathogenic T helper 17 cell generation by repressing T helper 2-biased programs
Article Snippet: For qRT-PCR analysis, total RNA was extracted from lymphocytes using TRizol reagent (Invitrogen) and reverse-transcribed into cDNA with Superscript III reverse transcriptase (Bio-Rad) per manufacturer’s protocols. .. RNA-seq libraries were generated and poly(A) enriched with 1 microgram of RNA as input using the TruSeq RNA Sample Prep Kit (Illumina, San Diego, CA).

Article Title: RAS P21 Protein Activator 3 (RASA3) specifically promotes pathogenic T helper 17 cell generation by repressing T helper 2-biased programs
Article Snippet: For qRT-PCR analysis, total RNA was extracted from lymphocytes using TRizol reagent (Invitrogen) and reverse-transcribed into cDNA with Superscript III reverse transcriptase (Bio-Rad) per manufacturer’s protocols. .. RNA-seq libraries were generated and poly(A) enriched with 1 microgram of RNA as input using the TruSeq RNA Sample Prep Kit (Illumina, San Diego, CA).

Synthesized:

Article Title: Collaborative interactions between type-2 innate lymphoid cells and Ag-specific CD4+Th2 cells exacerbate murine allergic airway diseases with prominent eosinophilia
Article Snippet: .. RNA from sorted cell populations was isolated with an RNeasy Plus Mini kit (Qiagen), and cDNA templates were synthesized with SuperScript III reverse transcriptase (BioRad). .. Quantitative real-time PCR analyses were performed with SYBR Green Chemistry (Applied Biosystems) in an ABI Prism 7900 detection system using previously described primer sets ( , ).

Article Title: Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W]Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W] [OA]
Article Snippet: .. For P. abies , total RNA was isolated from needles or buds following the protocol described by with minor modifications, while for Arabidopsis, the Qiagen RNeasy Mini kit was used in accordance with the manufacturer’s recommendations. cDNAs were synthesized from 0.5 μg of total RNA using SuperScript III reverse transcriptase and random hexamer primers. qRT-PCR was performed according to or on a MyiQ Real-Time PCR Detection system (Bio-Rad) with the thermal conditions 95°C for 7 min followed by 40 cycles of 95°C for 10 s and 60°C for 30 s. Each reaction was performed in duplicate containing 12 μL of DyNAmo Flash SYBRGreen (DyNAmo Flash SYBRGreen qPCR kit; Finnzymes), 0.5 m m of each primer, and 4.75 μL of cDNA (diluted 1:100) in a total volume of 23.75 μL. ..

Article Title: Kr?ppel-like factor 4 is induced by rapamycin and mediates the anti-proliferative effect of rapamycin in rat carotid arteries after balloon injury
Article Snippet: .. Total RNA was isolated from VSMCs and arterial segments with Trizol Reagent (Invitrogen, Grand Island, NY, USA). cDNA was synthesized with the use of Superscript III reverse transcriptase and oligo-(dT) primer. qPCR was performed with the iQ™ SYBR Green PCR Supermix in the DNA Engine Opticon real-time system (Bio-Rad, Hercules, CA, USA), with GAPDH used as an internal control. ..

Isolation:

Article Title: Collaborative interactions between type-2 innate lymphoid cells and Ag-specific CD4+Th2 cells exacerbate murine allergic airway diseases with prominent eosinophilia
Article Snippet: .. RNA from sorted cell populations was isolated with an RNeasy Plus Mini kit (Qiagen), and cDNA templates were synthesized with SuperScript III reverse transcriptase (BioRad). .. Quantitative real-time PCR analyses were performed with SYBR Green Chemistry (Applied Biosystems) in an ABI Prism 7900 detection system using previously described primer sets ( , ).

Article Title: Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W]Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W] [OA]
Article Snippet: .. For P. abies , total RNA was isolated from needles or buds following the protocol described by with minor modifications, while for Arabidopsis, the Qiagen RNeasy Mini kit was used in accordance with the manufacturer’s recommendations. cDNAs were synthesized from 0.5 μg of total RNA using SuperScript III reverse transcriptase and random hexamer primers. qRT-PCR was performed according to or on a MyiQ Real-Time PCR Detection system (Bio-Rad) with the thermal conditions 95°C for 7 min followed by 40 cycles of 95°C for 10 s and 60°C for 30 s. Each reaction was performed in duplicate containing 12 μL of DyNAmo Flash SYBRGreen (DyNAmo Flash SYBRGreen qPCR kit; Finnzymes), 0.5 m m of each primer, and 4.75 μL of cDNA (diluted 1:100) in a total volume of 23.75 μL. ..

Article Title: Endotrophin triggers adipose tissue fibrosis and metabolic dysfunction
Article Snippet: Total RNAs were extracted in Trizol (Invitrogen, Carlsbad, CA) using a TissueLyser (Qiagen, Valenica, CA) and then isolated with the RNeasy RNA extraction kit (Qiagen) following the company's protocol. .. The purity and quantity of the mRNAs were determined by absorbance at 260/280 nm. cDNAs were prepared by reverse transcribing 500 ng of total mRNAs with a kit containing Superscript III reverse transcriptase and oligo (dT)20 (Bio-Rad). qPCR reactions were carried out on an ABI Prism 7900 HT sequence detection system (Applied Biosystems).

Article Title: Kr?ppel-like factor 4 is induced by rapamycin and mediates the anti-proliferative effect of rapamycin in rat carotid arteries after balloon injury
Article Snippet: .. Total RNA was isolated from VSMCs and arterial segments with Trizol Reagent (Invitrogen, Grand Island, NY, USA). cDNA was synthesized with the use of Superscript III reverse transcriptase and oligo-(dT) primer. qPCR was performed with the iQ™ SYBR Green PCR Supermix in the DNA Engine Opticon real-time system (Bio-Rad, Hercules, CA, USA), with GAPDH used as an internal control. ..

Article Title: Mov10 suppresses retroelements and regulates neuronal development and function in the developing brain
Article Snippet: To isolate associated RNA, the IPs were treated with proteinase K followed by TRIzol (Ambion) extraction for RNA isolation. .. The RNA was extracted with phenol-chloroform and precipitated in ethanol and converted into cDNA using Oligo dT primer and SuperScript III Reverse Transcriptase. qRT-PCR was performed with iQ SYBR Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA) using a StepOnePlus RT PCR machine (Applied Biosystems) with gene-specific primers.

Magnetic Beads:

Article Title: The PPARδ Agonist GW501516 Improves Lipolytic/Lipogenic Balance through CPT1 and PEPCK during the Development of Pre-Implantation Bovine Embryos
Article Snippet: Three hundred microliters of washing buffer A were used to wash the magnetic beads harboring the hybridized mRNA and oligo (dT). .. To denature the secondary structures, bound mRNAs were resuspended in 8 μL of 10 mM Tris-HCl and heated at 65 °C for 5 min, followed by rapid quenching of the reaction on ice for 3 min. Superscript III reverse transcriptase was used for mRNA to reverse-transcribe into first-strand cDNA (Bio-Rad Laboratories Hercules, CA, USA cat. #1708891).

RNA Sequencing Assay:

Article Title: RAS P21 Protein Activator 3 (RASA3) specifically promotes pathogenic T helper 17 cell generation by repressing T helper 2-biased programs
Article Snippet: Paragraph title: Quantitative RT-PCR (qRT-PCR) and RNA-seq analysis ... For qRT-PCR analysis, total RNA was extracted from lymphocytes using TRizol reagent (Invitrogen) and reverse-transcribed into cDNA with Superscript III reverse transcriptase (Bio-Rad) per manufacturer’s protocols.

Article Title: Mov10 suppresses retroelements and regulates neuronal development and function in the developing brain
Article Snippet: Paragraph title: Brain IP, RT-PCR, and RNA sequencing ... The RNA was extracted with phenol-chloroform and precipitated in ethanol and converted into cDNA using Oligo dT primer and SuperScript III Reverse Transcriptase. qRT-PCR was performed with iQ SYBR Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA) using a StepOnePlus RT PCR machine (Applied Biosystems) with gene-specific primers.

Generated:

Article Title: RAS P21 Protein Activator 3 (RASA3) specifically promotes pathogenic T helper 17 cell generation by repressing T helper 2-biased programs
Article Snippet: For qRT-PCR analysis, total RNA was extracted from lymphocytes using TRizol reagent (Invitrogen) and reverse-transcribed into cDNA with Superscript III reverse transcriptase (Bio-Rad) per manufacturer’s protocols. .. RNA-seq libraries were generated and poly(A) enriched with 1 microgram of RNA as input using the TruSeq RNA Sample Prep Kit (Illumina, San Diego, CA).

Article Title: RAS P21 Protein Activator 3 (RASA3) specifically promotes pathogenic T helper 17 cell generation by repressing T helper 2-biased programs
Article Snippet: For qRT-PCR analysis, total RNA was extracted from lymphocytes using TRizol reagent (Invitrogen) and reverse-transcribed into cDNA with Superscript III reverse transcriptase (Bio-Rad) per manufacturer’s protocols. .. RNA-seq libraries were generated and poly(A) enriched with 1 microgram of RNA as input using the TruSeq RNA Sample Prep Kit (Illumina, San Diego, CA).

Quantitative RT-PCR:

Article Title: RAS P21 Protein Activator 3 (RASA3) specifically promotes pathogenic T helper 17 cell generation by repressing T helper 2-biased programs
Article Snippet: .. For qRT-PCR analysis, total RNA was extracted from lymphocytes using TRizol reagent (Invitrogen) and reverse-transcribed into cDNA with Superscript III reverse transcriptase (Bio-Rad) per manufacturer’s protocols. .. Quantitative PCR (qPCR) was performed on QuantStudio® 6 Flex Real-Time PCR Syst em (ThermoFisher Scientific).

Article Title: Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W]Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W] [OA]
Article Snippet: .. For P. abies , total RNA was isolated from needles or buds following the protocol described by with minor modifications, while for Arabidopsis, the Qiagen RNeasy Mini kit was used in accordance with the manufacturer’s recommendations. cDNAs were synthesized from 0.5 μg of total RNA using SuperScript III reverse transcriptase and random hexamer primers. qRT-PCR was performed according to or on a MyiQ Real-Time PCR Detection system (Bio-Rad) with the thermal conditions 95°C for 7 min followed by 40 cycles of 95°C for 10 s and 60°C for 30 s. Each reaction was performed in duplicate containing 12 μL of DyNAmo Flash SYBRGreen (DyNAmo Flash SYBRGreen qPCR kit; Finnzymes), 0.5 m m of each primer, and 4.75 μL of cDNA (diluted 1:100) in a total volume of 23.75 μL. ..

Article Title: BPTF is essential for T cell homeostasis and function
Article Snippet: Paragraph title: Quantitative RT-PCR ... Total RNA was extracted from T cells with TRIzol reagent according to the manufacturer’s instructions (Invitrogen) and was reverse-transcribed into c-DNA with Superscript III reverse transcriptase (Bio-Rad).

Article Title: Kr?ppel-like factor 4 is induced by rapamycin and mediates the anti-proliferative effect of rapamycin in rat carotid arteries after balloon injury
Article Snippet: Paragraph title: Quantitative reverse-transcriptase-PCR (qRT-PCR) ... Total RNA was isolated from VSMCs and arterial segments with Trizol Reagent (Invitrogen, Grand Island, NY, USA). cDNA was synthesized with the use of Superscript III reverse transcriptase and oligo-(dT) primer. qPCR was performed with the iQ™ SYBR Green PCR Supermix in the DNA Engine Opticon real-time system (Bio-Rad, Hercules, CA, USA), with GAPDH used as an internal control.

Article Title: RAS P21 Protein Activator 3 (RASA3) specifically promotes pathogenic T helper 17 cell generation by repressing T helper 2-biased programs
Article Snippet: .. For qRT-PCR analysis, total RNA was extracted from lymphocytes using TRizol reagent (Invitrogen) and reverse-transcribed into cDNA with Superscript III reverse transcriptase (Bio-Rad) per manufacturer’s protocols. .. Quantitative PCR (qPCR) was performed on QuantStudio® 6 Flex Real-Time PCR Syst em (ThermoFisher Scientific).

Article Title: Mov10 suppresses retroelements and regulates neuronal development and function in the developing brain
Article Snippet: .. The RNA was extracted with phenol-chloroform and precipitated in ethanol and converted into cDNA using Oligo dT primer and SuperScript III Reverse Transcriptase. qRT-PCR was performed with iQ SYBR Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA) using a StepOnePlus RT PCR machine (Applied Biosystems) with gene-specific primers. .. For the brain IP-RNA sequencing, WT brains were homogenized in the same manner as described above but were not UV cross-linked.

Mouse Assay:

Article Title: Collaborative interactions between type-2 innate lymphoid cells and Ag-specific CD4+Th2 cells exacerbate murine allergic airway diseases with prominent eosinophilia
Article Snippet: RNA from sorted cell populations was isolated with an RNeasy Plus Mini kit (Qiagen), and cDNA templates were synthesized with SuperScript III reverse transcriptase (BioRad). .. Expression levels of target genes were normalized to endogenous Gapdh transcript levels, and relative quantification of samples was compared to the expression level of indicated genes in naïve CD4+ T cells isolated from naïve BALB/c mice as the baseline.

SYBR Green Assay:

Article Title: Collaborative interactions between type-2 innate lymphoid cells and Ag-specific CD4+Th2 cells exacerbate murine allergic airway diseases with prominent eosinophilia
Article Snippet: RNA from sorted cell populations was isolated with an RNeasy Plus Mini kit (Qiagen), and cDNA templates were synthesized with SuperScript III reverse transcriptase (BioRad). .. Quantitative real-time PCR analyses were performed with SYBR Green Chemistry (Applied Biosystems) in an ABI Prism 7900 detection system using previously described primer sets ( , ).

Article Title: Kr?ppel-like factor 4 is induced by rapamycin and mediates the anti-proliferative effect of rapamycin in rat carotid arteries after balloon injury
Article Snippet: .. Total RNA was isolated from VSMCs and arterial segments with Trizol Reagent (Invitrogen, Grand Island, NY, USA). cDNA was synthesized with the use of Superscript III reverse transcriptase and oligo-(dT) primer. qPCR was performed with the iQ™ SYBR Green PCR Supermix in the DNA Engine Opticon real-time system (Bio-Rad, Hercules, CA, USA), with GAPDH used as an internal control. ..

Article Title: Mov10 suppresses retroelements and regulates neuronal development and function in the developing brain
Article Snippet: .. The RNA was extracted with phenol-chloroform and precipitated in ethanol and converted into cDNA using Oligo dT primer and SuperScript III Reverse Transcriptase. qRT-PCR was performed with iQ SYBR Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA) using a StepOnePlus RT PCR machine (Applied Biosystems) with gene-specific primers. .. For the brain IP-RNA sequencing, WT brains were homogenized in the same manner as described above but were not UV cross-linked.

Polymerase Chain Reaction:

Article Title: Kr?ppel-like factor 4 is induced by rapamycin and mediates the anti-proliferative effect of rapamycin in rat carotid arteries after balloon injury
Article Snippet: .. Total RNA was isolated from VSMCs and arterial segments with Trizol Reagent (Invitrogen, Grand Island, NY, USA). cDNA was synthesized with the use of Superscript III reverse transcriptase and oligo-(dT) primer. qPCR was performed with the iQ™ SYBR Green PCR Supermix in the DNA Engine Opticon real-time system (Bio-Rad, Hercules, CA, USA), with GAPDH used as an internal control. ..

Random Hexamer Labeling:

Article Title: Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W]Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W] [OA]
Article Snippet: .. For P. abies , total RNA was isolated from needles or buds following the protocol described by with minor modifications, while for Arabidopsis, the Qiagen RNeasy Mini kit was used in accordance with the manufacturer’s recommendations. cDNAs were synthesized from 0.5 μg of total RNA using SuperScript III reverse transcriptase and random hexamer primers. qRT-PCR was performed according to or on a MyiQ Real-Time PCR Detection system (Bio-Rad) with the thermal conditions 95°C for 7 min followed by 40 cycles of 95°C for 10 s and 60°C for 30 s. Each reaction was performed in duplicate containing 12 μL of DyNAmo Flash SYBRGreen (DyNAmo Flash SYBRGreen qPCR kit; Finnzymes), 0.5 m m of each primer, and 4.75 μL of cDNA (diluted 1:100) in a total volume of 23.75 μL. ..

Immunoprecipitation:

Article Title: Mov10 suppresses retroelements and regulates neuronal development and function in the developing brain
Article Snippet: Triturated cells were lysed in lysis buffer (50 mM Tris-Cl 7.5, 300 mM NaCl, 30 mM ethylenediaminetetraacetic acid (EDTA), 0.5% Triton), cleared by ultracentrifugation (35,000 rpm at 35 min at 4 °C), and sequentially immunoprecipitated with an irrelevant rabbit polyclonal antibody (anti-EGFP, Clontech, Mountain View, CA, USA; RRID: AB_10013427) followed by IP with Mov10 antibody (A301-571A, RRID: AB_1040002, Bethyl). .. The RNA was extracted with phenol-chloroform and precipitated in ethanol and converted into cDNA using Oligo dT primer and SuperScript III Reverse Transcriptase. qRT-PCR was performed with iQ SYBR Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA) using a StepOnePlus RT PCR machine (Applied Biosystems) with gene-specific primers.

Expressing:

Article Title: Collaborative interactions between type-2 innate lymphoid cells and Ag-specific CD4+Th2 cells exacerbate murine allergic airway diseases with prominent eosinophilia
Article Snippet: RNA from sorted cell populations was isolated with an RNeasy Plus Mini kit (Qiagen), and cDNA templates were synthesized with SuperScript III reverse transcriptase (BioRad). .. Expression levels of target genes were normalized to endogenous Gapdh transcript levels, and relative quantification of samples was compared to the expression level of indicated genes in naïve CD4+ T cells isolated from naïve BALB/c mice as the baseline.

Article Title: Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W]Evolution of the PEBP Gene Family in Plants: Functional Diversification in Seed Plant Evolution 1 [W] [OA]
Article Snippet: Paragraph title: Quantitative Gene Expression Analysis ... For P. abies , total RNA was isolated from needles or buds following the protocol described by with minor modifications, while for Arabidopsis, the Qiagen RNeasy Mini kit was used in accordance with the manufacturer’s recommendations. cDNAs were synthesized from 0.5 μg of total RNA using SuperScript III reverse transcriptase and random hexamer primers. qRT-PCR was performed according to or on a MyiQ Real-Time PCR Detection system (Bio-Rad) with the thermal conditions 95°C for 7 min followed by 40 cycles of 95°C for 10 s and 60°C for 30 s. Each reaction was performed in duplicate containing 12 μL of DyNAmo Flash SYBRGreen (DyNAmo Flash SYBRGreen qPCR kit; Finnzymes), 0.5 m m of each primer, and 4.75 μL of cDNA (diluted 1:100) in a total volume of 23.75 μL.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Mov10 suppresses retroelements and regulates neuronal development and function in the developing brain
Article Snippet: .. The RNA was extracted with phenol-chloroform and precipitated in ethanol and converted into cDNA using Oligo dT primer and SuperScript III Reverse Transcriptase. qRT-PCR was performed with iQ SYBR Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA) using a StepOnePlus RT PCR machine (Applied Biosystems) with gene-specific primers. .. For the brain IP-RNA sequencing, WT brains were homogenized in the same manner as described above but were not UV cross-linked.

Lysis:

Article Title: Mov10 suppresses retroelements and regulates neuronal development and function in the developing brain
Article Snippet: Both IPs were washed sequentially for 10 min with lysis buffer and twice with wash buffer (1× PBS, 0.1% sodium dodecyl sulfate (SDS), 0.5% sodium deoxycholate, 1% NP40). .. The RNA was extracted with phenol-chloroform and precipitated in ethanol and converted into cDNA using Oligo dT primer and SuperScript III Reverse Transcriptase. qRT-PCR was performed with iQ SYBR Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA) using a StepOnePlus RT PCR machine (Applied Biosystems) with gene-specific primers.

Article Title: The PPARδ Agonist GW501516 Improves Lipolytic/Lipogenic Balance through CPT1 and PEPCK during the Development of Pre-Implantation Bovine Embryos
Article Snippet: In 100 μL of lysis buffer, oocytes or blastocysts were suspended and vortexed at room temperature for 2 min. .. To denature the secondary structures, bound mRNAs were resuspended in 8 μL of 10 mM Tris-HCl and heated at 65 °C for 5 min, followed by rapid quenching of the reaction on ice for 3 min. Superscript III reverse transcriptase was used for mRNA to reverse-transcribe into first-strand cDNA (Bio-Rad Laboratories Hercules, CA, USA cat. #1708891).

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    Bio-Rad superscript iii reverse transcriptase
    PPARs’ <t>mRNA</t> and protein expression during maturation and embryo development. ( A ) RT-PCR-based PPARα, PPARγ, and PPARδ expression at the COCs (cumulus oocyte complexes) (10 per sample), MII oocyte (20 per sample), two-cell embryo (20 per sample), 3.5-day embryo (10 per sample) and day-8 blastocyst stages (five per sample). ( B ) Relative mRNA expressions of PPARα, PPARγ and PPARδ from the COCs stage to day-8 blastocysts. ( C ) Development stage-dependent immunofluorescent expression of PPARα and PPARγ ( n = 15 per sample). ( D ) Immunofluorescent co-localization of PPARδ and PEPCK from COCs to day-8 blastocysts ( n = 15 per sample). The experiments were repeated <t>three</t> times, and the data are shown here as mean ± S.E.M. N.S., not significant. Significance: * p
    Superscript Iii Reverse Transcriptase, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript iii reverse transcriptase/product/Bio-Rad
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    95
    Bio-Rad superscript iii reverse transcriptase enzyme
    PPARs’ <t>mRNA</t> and protein expression during maturation and embryo development. ( A ) RT-PCR-based PPARα, PPARγ, and PPARδ expression at the COCs (cumulus oocyte complexes) (10 per sample), MII oocyte (20 per sample), two-cell embryo (20 per sample), 3.5-day embryo (10 per sample) and day-8 blastocyst stages (five per sample). ( B ) Relative mRNA expressions of PPARα, PPARγ and PPARδ from the COCs stage to day-8 blastocysts. ( C ) Development stage-dependent immunofluorescent expression of PPARα and PPARγ ( n = 15 per sample). ( D ) Immunofluorescent co-localization of PPARδ and PEPCK from COCs to day-8 blastocysts ( n = 15 per sample). The experiments were repeated <t>three</t> times, and the data are shown here as mean ± S.E.M. N.S., not significant. Significance: * p
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    PPARs’ mRNA and protein expression during maturation and embryo development. ( A ) RT-PCR-based PPARα, PPARγ, and PPARδ expression at the COCs (cumulus oocyte complexes) (10 per sample), MII oocyte (20 per sample), two-cell embryo (20 per sample), 3.5-day embryo (10 per sample) and day-8 blastocyst stages (five per sample). ( B ) Relative mRNA expressions of PPARα, PPARγ and PPARδ from the COCs stage to day-8 blastocysts. ( C ) Development stage-dependent immunofluorescent expression of PPARα and PPARγ ( n = 15 per sample). ( D ) Immunofluorescent co-localization of PPARδ and PEPCK from COCs to day-8 blastocysts ( n = 15 per sample). The experiments were repeated three times, and the data are shown here as mean ± S.E.M. N.S., not significant. Significance: * p

    Journal: International Journal of Molecular Sciences

    Article Title: The PPARδ Agonist GW501516 Improves Lipolytic/Lipogenic Balance through CPT1 and PEPCK during the Development of Pre-Implantation Bovine Embryos

    doi: 10.3390/ijms20236066

    Figure Lengend Snippet: PPARs’ mRNA and protein expression during maturation and embryo development. ( A ) RT-PCR-based PPARα, PPARγ, and PPARδ expression at the COCs (cumulus oocyte complexes) (10 per sample), MII oocyte (20 per sample), two-cell embryo (20 per sample), 3.5-day embryo (10 per sample) and day-8 blastocyst stages (five per sample). ( B ) Relative mRNA expressions of PPARα, PPARγ and PPARδ from the COCs stage to day-8 blastocysts. ( C ) Development stage-dependent immunofluorescent expression of PPARα and PPARγ ( n = 15 per sample). ( D ) Immunofluorescent co-localization of PPARδ and PEPCK from COCs to day-8 blastocysts ( n = 15 per sample). The experiments were repeated three times, and the data are shown here as mean ± S.E.M. N.S., not significant. Significance: * p

    Article Snippet: To denature the secondary structures, bound mRNAs were resuspended in 8 μL of 10 mM Tris-HCl and heated at 65 °C for 5 min, followed by rapid quenching of the reaction on ice for 3 min. Superscript III reverse transcriptase was used for mRNA to reverse-transcribe into first-strand cDNA (Bio-Rad Laboratories Hercules, CA, USA cat. #1708891).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    PPARδ reversibility and lipid metabolism in bovine embryos. ( A ) Immunofluorescent co-localization of PEPCK and CPT1 in control-, GSK3787-, and GW501516 IVC-treated day-8 blastocysts (15 per group). The original magnification is ×200. ( B ) RT-qPCR-based mRNA quantification of ATGL, LMF1, LMF2, and LPL in day-8 blastocysts (five per sample). ( C ) Western blot analysis of SIRT1 and p-mTOR in control-, GSK3787-, and GW501516 IVC-treated blastocysts (20 per group). The experiments were repeated three times, and the data are shown here as mean ± S.E.M. * p

    Journal: International Journal of Molecular Sciences

    Article Title: The PPARδ Agonist GW501516 Improves Lipolytic/Lipogenic Balance through CPT1 and PEPCK during the Development of Pre-Implantation Bovine Embryos

    doi: 10.3390/ijms20236066

    Figure Lengend Snippet: PPARδ reversibility and lipid metabolism in bovine embryos. ( A ) Immunofluorescent co-localization of PEPCK and CPT1 in control-, GSK3787-, and GW501516 IVC-treated day-8 blastocysts (15 per group). The original magnification is ×200. ( B ) RT-qPCR-based mRNA quantification of ATGL, LMF1, LMF2, and LPL in day-8 blastocysts (five per sample). ( C ) Western blot analysis of SIRT1 and p-mTOR in control-, GSK3787-, and GW501516 IVC-treated blastocysts (20 per group). The experiments were repeated three times, and the data are shown here as mean ± S.E.M. * p

    Article Snippet: To denature the secondary structures, bound mRNAs were resuspended in 8 μL of 10 mM Tris-HCl and heated at 65 °C for 5 min, followed by rapid quenching of the reaction on ice for 3 min. Superscript III reverse transcriptase was used for mRNA to reverse-transcribe into first-strand cDNA (Bio-Rad Laboratories Hercules, CA, USA cat. #1708891).

    Techniques: Quantitative RT-PCR, Western Blot