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Roche superscript iii reverse transcriptase
Superscript Iii Reverse Transcriptase, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/superscript iii reverse transcriptase/product/Roche
Average 94 stars, based on 6 article reviews
Price from $9.99 to $1999.99
superscript iii reverse transcriptase - by Bioz Stars, 2020-04
94/100 stars

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Nucleic Acid Electrophoresis:

Article Title: Meiotic arrest with roscovitine and follicular fluid improves cytoplasmic maturation of porcine oocytes by promoting chromatin de-condensation and gene transcription
Article Snippet: Thus, 2 µl of each RNA sample was mixed with 1 µl of Oligo(dT)18 (Fermentas) and 10 µl of DEPC-dH2 O in a 0.2 ml reaction tube, and the mixture was incubated in a PCR instrument at 65 °C for 10 min. At the end of the incubation period, the reaction tube was cooled on ice for 2 min, and then, 4 µl of 5× reverse transcriptase (RT) buffer, 2 µl of deoxynucleotide (dNTP), 0.5 µl of RNase inhibitor, and 0.5 µl of Superscript III Reverse Transcriptase (Roche) were added to the reaction tube. .. The cycle amplification conditions: (1) an initial denaturation step at 95 °C for 3 min; (2) 40 cycles at 95 °C for 20 sec; and (3) annealing temperature for 20 sec. To determine specificity of the reaction, PCR products were analyzed by sequencing, dissociation curve analysis, and gel electrophoresis.

Transfection:

Article Title: ACBD3 Interaction with TBC1 Domain 22 Protein Is Differentially Affected by Enteroviral and Kobuviral 3A Protein Binding
Article Snippet: For qRT-PCR, 1 µg of Aichi virus 3A-Flag or poliovirus 3A-Flag was transfected into a 6-well plate of HEK293T cells in log phase and harvested 48 h later. .. Two micrograms of total RNA from HEK293T cells was reverse transcribed using SuperScript III reverse transcriptase and oligo(dT)20 , and qRT-PCR was performed using the 480 DNA SYBR Green I master mix (Roche) on a LightCycler (Roche).

Amplification:

Article Title: Meiotic arrest with roscovitine and follicular fluid improves cytoplasmic maturation of porcine oocytes by promoting chromatin de-condensation and gene transcription
Article Snippet: Thus, 2 µl of each RNA sample was mixed with 1 µl of Oligo(dT)18 (Fermentas) and 10 µl of DEPC-dH2 O in a 0.2 ml reaction tube, and the mixture was incubated in a PCR instrument at 65 °C for 10 min. At the end of the incubation period, the reaction tube was cooled on ice for 2 min, and then, 4 µl of 5× reverse transcriptase (RT) buffer, 2 µl of deoxynucleotide (dNTP), 0.5 µl of RNase inhibitor, and 0.5 µl of Superscript III Reverse Transcriptase (Roche) were added to the reaction tube. .. A Mx3005 P Real-Time PCR System (Stratagene) was used for mRNA quantification. a 10 µl reaction volume was used for the amplification reactions including 1 µl of cDNA, 5 µl of 2× SYBR Green Master Mix (Agilent), 0.15 µl of 500-fold diluted reference dye, 3.25 µl of RNase-free water, and 0.3 µl each of forward and reverse gene-specific primers (10 μM).

Synthesized:

Article Title: Gadd45a sensitizes medulloblastoma cells to irradiation and suppresses MMP-9-mediated EMT
Article Snippet: .. With 1 μg of RNA used as a template, first-strand cDNA was synthesized using Superscript III reverse transcriptase (Roche Applied Science). .. PCR analysis using 100 ng of first-strand cDNA was completed using specific primers (Table ).

Article Title: Nucleolar Relocalization of RBM14 by Influenza A Virus NS1 Protein
Article Snippet: Cellular RNA was extracted using an RNeasy minikit (Invitrogen) following the manufacturer’s directions. cDNA corresponding to the viral mRNA of segment 6 (NA) was synthesized using tagged primers, thereby adding an 18-to-20-nucleotide tag at the 5′ end as follows: CCAGATCGTTCGAGTCGT . .. A 12-μl volume of preheated reaction mixture (10 μl First Strand buffer [Invitrogen; 2×] and 2 μl Superscript III reverse transcriptase) was added and incubated for 1 h. Quantitative PCR (qPCR) was performed with a SYBR green qPCR kit (Roche; catalog no. 507203180) using a LightCycler 480 system (Roche).

Isolation:

Article Title: Gadd45a sensitizes medulloblastoma cells to irradiation and suppresses MMP-9-mediated EMT
Article Snippet: Total RNA was isolated from control and treated cells using TRIZOL reagent (Invitrogen) according to the standard protocol. .. With 1 μg of RNA used as a template, first-strand cDNA was synthesized using Superscript III reverse transcriptase (Roche Applied Science).

Article Title: Oral AGE restriction ameliorates insulin resistance in obese individuals with the metabolic syndrome: a randomised controlled trial
Article Snippet: Paragraph title: RNA isolation and qRT-PCR ... First-strand cDNA synthesis was performed using SuperScript III Reverse Transcriptase (Roche, Indianapolis, IN, USA).

Article Title: Meiotic arrest with roscovitine and follicular fluid improves cytoplasmic maturation of porcine oocytes by promoting chromatin de-condensation and gene transcription
Article Snippet: Real-time RT-PCR RNA was extracted from 300 cumulus-free oocytes or the cumulus cells removed from 150 oocytes using a commercial RNA isolation kit (RNAqueous-Micro Kit, cat. no. AM1931). .. Thus, 2 µl of each RNA sample was mixed with 1 µl of Oligo(dT)18 (Fermentas) and 10 µl of DEPC-dH2 O in a 0.2 ml reaction tube, and the mixture was incubated in a PCR instrument at 65 °C for 10 min. At the end of the incubation period, the reaction tube was cooled on ice for 2 min, and then, 4 µl of 5× reverse transcriptase (RT) buffer, 2 µl of deoxynucleotide (dNTP), 0.5 µl of RNase inhibitor, and 0.5 µl of Superscript III Reverse Transcriptase (Roche) were added to the reaction tube.

Article Title: Hierarchical patterning modes orchestrate hair follicle morphogenesis
Article Snippet: .. qRT-PCR Total RNA was isolated from skin explants using Tri Reagent (Sigma) and treated with RQ1 DNAse (Promega) to remove contaminating genomic DNA. cDNA was synthesised from total RNA using random primers and Superscript III reverse transcriptase (Roche) in a 20-μl reaction. ..

Size-exclusion Chromatography:

Article Title: Meiotic arrest with roscovitine and follicular fluid improves cytoplasmic maturation of porcine oocytes by promoting chromatin de-condensation and gene transcription
Article Snippet: Thus, 2 µl of each RNA sample was mixed with 1 µl of Oligo(dT)18 (Fermentas) and 10 µl of DEPC-dH2 O in a 0.2 ml reaction tube, and the mixture was incubated in a PCR instrument at 65 °C for 10 min. At the end of the incubation period, the reaction tube was cooled on ice for 2 min, and then, 4 µl of 5× reverse transcriptase (RT) buffer, 2 µl of deoxynucleotide (dNTP), 0.5 µl of RNase inhibitor, and 0.5 µl of Superscript III Reverse Transcriptase (Roche) were added to the reaction tube. .. The cycle amplification conditions: (1) an initial denaturation step at 95 °C for 3 min; (2) 40 cycles at 95 °C for 20 sec; and (3) annealing temperature for 20 sec. To determine specificity of the reaction, PCR products were analyzed by sequencing, dissociation curve analysis, and gel electrophoresis.

Quantitative RT-PCR:

Article Title: Retinoic acid-independent expression of Meis2 during autopod patterning in the developing bat and mouse limb
Article Snippet: Relative qPCR analysis RT-qPCR was performed following the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) conventions [ ]. .. Experiments were performed using Amino allyl aRNA samples that were reverse transcribed into cDNA using Superscript III Reverse transcriptase (Roche Molecular Diagnostics, Pleasanton, CA, US) and second-round primers (Ambion, Austin, TX, USA).

Article Title: ACBD3 Interaction with TBC1 Domain 22 Protein Is Differentially Affected by Enteroviral and Kobuviral 3A Protein Binding
Article Snippet: .. Two micrograms of total RNA from HEK293T cells was reverse transcribed using SuperScript III reverse transcriptase and oligo(dT)20 , and qRT-PCR was performed using the 480 DNA SYBR Green I master mix (Roche) on a LightCycler (Roche). ..

Article Title: Dopaminergic neuron-specific deletion of p53 gene attenuates methamphetamine neurotoxicity
Article Snippet: .. Specifically, 1ug of total RNA was treated with RQ-1 RNase-free DNase I and was reverse transcribed into cDNA by Superscript III reverse transcriptase (Life Sciences) using random hexamers. cDNA levels for HPRT1 (hypoxanthine phosphoribosyltransferase 1), HMBS (hydroxymethylbilane synthase), and various target genes were acquired by quantitative RT-PCR using specific universal probe library primer probe sets (Roche) and a Roche Light Cycler II 480. .. Relative expression levels were calculated using the double delta Ct analysis method, which were compared to HMBS as a reference gene (n=8–10 for each group), as previously described ( ).

Article Title: Dopaminergic neuron-specific deletion of p53 gene is neuroprotective in an experimental Parkinson's disease model
Article Snippet: .. Total RNA (0.7 μg) was treated with RQ-1 Rnase-free Dnase I and reverse transcribed into cDNA using random hexamers by Superscript III reverse transcriptase (Life Sciences). cDNA levels for HPRT1 (hypoxanthine phosphoribosyltransferase 1), Hmbs (hydroxymethylbilane synthase) and various target genes were determined using specific universal probe Library primer probe sets (Roche) by quantitative RT-PCR using a Roche Light Cycler II 480. .. A relative expression level was calculated using the delta delta Ct method compared to Hmbs as a reference gene, with a N=7-8 for each group.

Article Title: Effect of the sonic hedgehog receptor smoothened on the survival and function of dopaminergic neurons
Article Snippet: .. Total RNA (1 ug) was treated with RQ-1 Rnase-free Dnase I and reverse transcribed into cDNA using random hexamers by Superscript III reverse transcriptase (Life Sciences). cDNA levels for HPRT1 (hypoxanthine phosphoribosyltransferase 1), Hmbs (hydroxymethylbilane synthase) and various target genes were determined, using specific universal probe Library primer probe sets (Roche), by quantitative RT-PCR using a Roche Light Cycler II 480. .. A relative expression level was calculated using the delta Ct method compared to Hprt1 as a reference gene, with n=7-8 for each group.

Article Title: Oral AGE restriction ameliorates insulin resistance in obese individuals with the metabolic syndrome: a randomised controlled trial
Article Snippet: Paragraph title: RNA isolation and qRT-PCR ... First-strand cDNA synthesis was performed using SuperScript III Reverse Transcriptase (Roche, Indianapolis, IN, USA).

Article Title: Meiotic arrest with roscovitine and follicular fluid improves cytoplasmic maturation of porcine oocytes by promoting chromatin de-condensation and gene transcription
Article Snippet: Paragraph title: Real-time RT-PCR ... Thus, 2 µl of each RNA sample was mixed with 1 µl of Oligo(dT)18 (Fermentas) and 10 µl of DEPC-dH2 O in a 0.2 ml reaction tube, and the mixture was incubated in a PCR instrument at 65 °C for 10 min. At the end of the incubation period, the reaction tube was cooled on ice for 2 min, and then, 4 µl of 5× reverse transcriptase (RT) buffer, 2 µl of deoxynucleotide (dNTP), 0.5 µl of RNase inhibitor, and 0.5 µl of Superscript III Reverse Transcriptase (Roche) were added to the reaction tube.

Article Title: Hierarchical patterning modes orchestrate hair follicle morphogenesis
Article Snippet: .. qRT-PCR Total RNA was isolated from skin explants using Tri Reagent (Sigma) and treated with RQ1 DNAse (Promega) to remove contaminating genomic DNA. cDNA was synthesised from total RNA using random primers and Superscript III reverse transcriptase (Roche) in a 20-μl reaction. ..

Purification:

Article Title: A Rho-actin signaling pathway shapes cell wall boundaries in Arabidopsis xylem vessels
Article Snippet: Quantitative reverse transcription PCR analysis Total RNA was prepared from Arabidopsis cells using the SDS-phenol method, treated with DNase, and purified using an RNeasy Plant Mini kit (QIAGEN). .. After reverse transcription with oligo(dT)20 priming and SuperScript III reverse transcriptase, quantitative reverse transcription PCR was performed using a LightCycler 96 Instrument (Roche Diagnostics) with FastStart Essential DNA Green Master (Roche Diagnostics).

SYBR Green Assay:

Article Title: ACBD3 Interaction with TBC1 Domain 22 Protein Is Differentially Affected by Enteroviral and Kobuviral 3A Protein Binding
Article Snippet: .. Two micrograms of total RNA from HEK293T cells was reverse transcribed using SuperScript III reverse transcriptase and oligo(dT)20 , and qRT-PCR was performed using the 480 DNA SYBR Green I master mix (Roche) on a LightCycler (Roche). ..

Article Title: Nucleolar Relocalization of RBM14 by Influenza A Virus NS1 Protein
Article Snippet: .. A 12-μl volume of preheated reaction mixture (10 μl First Strand buffer [Invitrogen; 2×] and 2 μl Superscript III reverse transcriptase) was added and incubated for 1 h. Quantitative PCR (qPCR) was performed with a SYBR green qPCR kit (Roche; catalog no. 507203180) using a LightCycler 480 system (Roche). .. A 3-μl volume of a 10-fold dilution of the cDNA was added to the qPCR reaction mixture (5 μl SYBR green qPCR mix, 1 μl of 2 μM forward primer, and 1 μl of 2 μM reverse primer).

Article Title: Oral AGE restriction ameliorates insulin resistance in obese individuals with the metabolic syndrome: a randomised controlled trial
Article Snippet: First-strand cDNA synthesis was performed using SuperScript III Reverse Transcriptase (Roche, Indianapolis, IN, USA). .. AGER1 (also known as DDOST ), receptor for AGEs ( RAGE , also known as AGER ) and SIRT1 mRNA expression were analysed by quantitative SYBR Green real-time PCR.

Article Title: Meiotic arrest with roscovitine and follicular fluid improves cytoplasmic maturation of porcine oocytes by promoting chromatin de-condensation and gene transcription
Article Snippet: Thus, 2 µl of each RNA sample was mixed with 1 µl of Oligo(dT)18 (Fermentas) and 10 µl of DEPC-dH2 O in a 0.2 ml reaction tube, and the mixture was incubated in a PCR instrument at 65 °C for 10 min. At the end of the incubation period, the reaction tube was cooled on ice for 2 min, and then, 4 µl of 5× reverse transcriptase (RT) buffer, 2 µl of deoxynucleotide (dNTP), 0.5 µl of RNase inhibitor, and 0.5 µl of Superscript III Reverse Transcriptase (Roche) were added to the reaction tube. .. A Mx3005 P Real-Time PCR System (Stratagene) was used for mRNA quantification. a 10 µl reaction volume was used for the amplification reactions including 1 µl of cDNA, 5 µl of 2× SYBR Green Master Mix (Agilent), 0.15 µl of 500-fold diluted reference dye, 3.25 µl of RNase-free water, and 0.3 µl each of forward and reverse gene-specific primers (10 μM).

Article Title: Hierarchical patterning modes orchestrate hair follicle morphogenesis
Article Snippet: qRT-PCR Total RNA was isolated from skin explants using Tri Reagent (Sigma) and treated with RQ1 DNAse (Promega) to remove contaminating genomic DNA. cDNA was synthesised from total RNA using random primers and Superscript III reverse transcriptase (Roche) in a 20-μl reaction. .. Each reaction was performed in a 20-μl volume using Universal SYBR Green Master Mix (Roche) containing Rox reference dye.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Gadd45a sensitizes medulloblastoma cells to irradiation and suppresses MMP-9-mediated EMT
Article Snippet: Paragraph title: Reverse-Transcription Polymerase Chain Reaction (PCR) ... With 1 μg of RNA used as a template, first-strand cDNA was synthesized using Superscript III reverse transcriptase (Roche Applied Science).

Incubation:

Article Title: Nucleolar Relocalization of RBM14 by Influenza A Virus NS1 Protein
Article Snippet: .. A 12-μl volume of preheated reaction mixture (10 μl First Strand buffer [Invitrogen; 2×] and 2 μl Superscript III reverse transcriptase) was added and incubated for 1 h. Quantitative PCR (qPCR) was performed with a SYBR green qPCR kit (Roche; catalog no. 507203180) using a LightCycler 480 system (Roche). .. A 3-μl volume of a 10-fold dilution of the cDNA was added to the qPCR reaction mixture (5 μl SYBR green qPCR mix, 1 μl of 2 μM forward primer, and 1 μl of 2 μM reverse primer).

Article Title: Meiotic arrest with roscovitine and follicular fluid improves cytoplasmic maturation of porcine oocytes by promoting chromatin de-condensation and gene transcription
Article Snippet: .. Thus, 2 µl of each RNA sample was mixed with 1 µl of Oligo(dT)18 (Fermentas) and 10 µl of DEPC-dH2 O in a 0.2 ml reaction tube, and the mixture was incubated in a PCR instrument at 65 °C for 10 min. At the end of the incubation period, the reaction tube was cooled on ice for 2 min, and then, 4 µl of 5× reverse transcriptase (RT) buffer, 2 µl of deoxynucleotide (dNTP), 0.5 µl of RNase inhibitor, and 0.5 µl of Superscript III Reverse Transcriptase (Roche) were added to the reaction tube. ..

Polymerase Chain Reaction:

Article Title: A Rho-actin signaling pathway shapes cell wall boundaries in Arabidopsis xylem vessels
Article Snippet: .. After reverse transcription with oligo(dT)20 priming and SuperScript III reverse transcriptase, quantitative reverse transcription PCR was performed using a LightCycler 96 Instrument (Roche Diagnostics) with FastStart Essential DNA Green Master (Roche Diagnostics). .. UBQ10 expression was used as a control.

Article Title: Gadd45a sensitizes medulloblastoma cells to irradiation and suppresses MMP-9-mediated EMT
Article Snippet: Paragraph title: Reverse-Transcription Polymerase Chain Reaction (PCR) ... With 1 μg of RNA used as a template, first-strand cDNA was synthesized using Superscript III reverse transcriptase (Roche Applied Science).

Article Title: Meiotic arrest with roscovitine and follicular fluid improves cytoplasmic maturation of porcine oocytes by promoting chromatin de-condensation and gene transcription
Article Snippet: .. Thus, 2 µl of each RNA sample was mixed with 1 µl of Oligo(dT)18 (Fermentas) and 10 µl of DEPC-dH2 O in a 0.2 ml reaction tube, and the mixture was incubated in a PCR instrument at 65 °C for 10 min. At the end of the incubation period, the reaction tube was cooled on ice for 2 min, and then, 4 µl of 5× reverse transcriptase (RT) buffer, 2 µl of deoxynucleotide (dNTP), 0.5 µl of RNase inhibitor, and 0.5 µl of Superscript III Reverse Transcriptase (Roche) were added to the reaction tube. ..

Labeling:

Article Title: Effect of the sonic hedgehog receptor smoothened on the survival and function of dopaminergic neurons
Article Snippet: Total RNA (1 ug) was treated with RQ-1 Rnase-free Dnase I and reverse transcribed into cDNA using random hexamers by Superscript III reverse transcriptase (Life Sciences). cDNA levels for HPRT1 (hypoxanthine phosphoribosyltransferase 1), Hmbs (hydroxymethylbilane synthase) and various target genes were determined, using specific universal probe Library primer probe sets (Roche), by quantitative RT-PCR using a Roche Light Cycler II 480. .. Primers and 6-carboxyfluorescein (FAM) labeled probes used in the quantitative RT-PCR for each gene are listed in .

Expressing:

Article Title: A Rho-actin signaling pathway shapes cell wall boundaries in Arabidopsis xylem vessels
Article Snippet: After reverse transcription with oligo(dT)20 priming and SuperScript III reverse transcriptase, quantitative reverse transcription PCR was performed using a LightCycler 96 Instrument (Roche Diagnostics) with FastStart Essential DNA Green Master (Roche Diagnostics). .. UBQ10 expression was used as a control.

Article Title: Dopaminergic neuron-specific deletion of p53 gene attenuates methamphetamine neurotoxicity
Article Snippet: Specifically, 1ug of total RNA was treated with RQ-1 RNase-free DNase I and was reverse transcribed into cDNA by Superscript III reverse transcriptase (Life Sciences) using random hexamers. cDNA levels for HPRT1 (hypoxanthine phosphoribosyltransferase 1), HMBS (hydroxymethylbilane synthase), and various target genes were acquired by quantitative RT-PCR using specific universal probe library primer probe sets (Roche) and a Roche Light Cycler II 480. .. Relative expression levels were calculated using the double delta Ct analysis method, which were compared to HMBS as a reference gene (n=8–10 for each group), as previously described ( ).

Article Title: Dopaminergic neuron-specific deletion of p53 gene is neuroprotective in an experimental Parkinson's disease model
Article Snippet: Total RNA (0.7 μg) was treated with RQ-1 Rnase-free Dnase I and reverse transcribed into cDNA using random hexamers by Superscript III reverse transcriptase (Life Sciences). cDNA levels for HPRT1 (hypoxanthine phosphoribosyltransferase 1), Hmbs (hydroxymethylbilane synthase) and various target genes were determined using specific universal probe Library primer probe sets (Roche) by quantitative RT-PCR using a Roche Light Cycler II 480. .. A relative expression level was calculated using the delta delta Ct method compared to Hmbs as a reference gene, with a N=7-8 for each group.

Article Title: Effect of the sonic hedgehog receptor smoothened on the survival and function of dopaminergic neurons
Article Snippet: Total RNA (1 ug) was treated with RQ-1 Rnase-free Dnase I and reverse transcribed into cDNA using random hexamers by Superscript III reverse transcriptase (Life Sciences). cDNA levels for HPRT1 (hypoxanthine phosphoribosyltransferase 1), Hmbs (hydroxymethylbilane synthase) and various target genes were determined, using specific universal probe Library primer probe sets (Roche), by quantitative RT-PCR using a Roche Light Cycler II 480. .. A relative expression level was calculated using the delta Ct method compared to Hprt1 as a reference gene, with n=7-8 for each group.

Article Title: Oral AGE restriction ameliorates insulin resistance in obese individuals with the metabolic syndrome: a randomised controlled trial
Article Snippet: First-strand cDNA synthesis was performed using SuperScript III Reverse Transcriptase (Roche, Indianapolis, IN, USA). .. AGER1 (also known as DDOST ), receptor for AGEs ( RAGE , also known as AGER ) and SIRT1 mRNA expression were analysed by quantitative SYBR Green real-time PCR.

Sequencing:

Article Title: Meiotic arrest with roscovitine and follicular fluid improves cytoplasmic maturation of porcine oocytes by promoting chromatin de-condensation and gene transcription
Article Snippet: Thus, 2 µl of each RNA sample was mixed with 1 µl of Oligo(dT)18 (Fermentas) and 10 µl of DEPC-dH2 O in a 0.2 ml reaction tube, and the mixture was incubated in a PCR instrument at 65 °C for 10 min. At the end of the incubation period, the reaction tube was cooled on ice for 2 min, and then, 4 µl of 5× reverse transcriptase (RT) buffer, 2 µl of deoxynucleotide (dNTP), 0.5 µl of RNase inhibitor, and 0.5 µl of Superscript III Reverse Transcriptase (Roche) were added to the reaction tube. .. The cycle amplification conditions: (1) an initial denaturation step at 95 °C for 3 min; (2) 40 cycles at 95 °C for 20 sec; and (3) annealing temperature for 20 sec. To determine specificity of the reaction, PCR products were analyzed by sequencing, dissociation curve analysis, and gel electrophoresis.

Real-time Polymerase Chain Reaction:

Article Title: Retinoic acid-independent expression of Meis2 during autopod patterning in the developing bat and mouse limb
Article Snippet: Paragraph title: Relative qPCR analysis ... Experiments were performed using Amino allyl aRNA samples that were reverse transcribed into cDNA using Superscript III Reverse transcriptase (Roche Molecular Diagnostics, Pleasanton, CA, US) and second-round primers (Ambion, Austin, TX, USA).

Article Title: Nucleolar Relocalization of RBM14 by Influenza A Virus NS1 Protein
Article Snippet: .. A 12-μl volume of preheated reaction mixture (10 μl First Strand buffer [Invitrogen; 2×] and 2 μl Superscript III reverse transcriptase) was added and incubated for 1 h. Quantitative PCR (qPCR) was performed with a SYBR green qPCR kit (Roche; catalog no. 507203180) using a LightCycler 480 system (Roche). .. A 3-μl volume of a 10-fold dilution of the cDNA was added to the qPCR reaction mixture (5 μl SYBR green qPCR mix, 1 μl of 2 μM forward primer, and 1 μl of 2 μM reverse primer).

Article Title: Oral AGE restriction ameliorates insulin resistance in obese individuals with the metabolic syndrome: a randomised controlled trial
Article Snippet: First-strand cDNA synthesis was performed using SuperScript III Reverse Transcriptase (Roche, Indianapolis, IN, USA). .. AGER1 (also known as DDOST ), receptor for AGEs ( RAGE , also known as AGER ) and SIRT1 mRNA expression were analysed by quantitative SYBR Green real-time PCR.

Article Title: Meiotic arrest with roscovitine and follicular fluid improves cytoplasmic maturation of porcine oocytes by promoting chromatin de-condensation and gene transcription
Article Snippet: Thus, 2 µl of each RNA sample was mixed with 1 µl of Oligo(dT)18 (Fermentas) and 10 µl of DEPC-dH2 O in a 0.2 ml reaction tube, and the mixture was incubated in a PCR instrument at 65 °C for 10 min. At the end of the incubation period, the reaction tube was cooled on ice for 2 min, and then, 4 µl of 5× reverse transcriptase (RT) buffer, 2 µl of deoxynucleotide (dNTP), 0.5 µl of RNase inhibitor, and 0.5 µl of Superscript III Reverse Transcriptase (Roche) were added to the reaction tube. .. A Mx3005 P Real-Time PCR System (Stratagene) was used for mRNA quantification. a 10 µl reaction volume was used for the amplification reactions including 1 µl of cDNA, 5 µl of 2× SYBR Green Master Mix (Agilent), 0.15 µl of 500-fold diluted reference dye, 3.25 µl of RNase-free water, and 0.3 µl each of forward and reverse gene-specific primers (10 μM).

Mouse Assay:

Article Title: Dopaminergic neuron-specific deletion of p53 gene attenuates methamphetamine neurotoxicity
Article Snippet: After euthanization, brains were immediately harvested from the mice and chilled on ice. .. Specifically, 1ug of total RNA was treated with RQ-1 RNase-free DNase I and was reverse transcribed into cDNA by Superscript III reverse transcriptase (Life Sciences) using random hexamers. cDNA levels for HPRT1 (hypoxanthine phosphoribosyltransferase 1), HMBS (hydroxymethylbilane synthase), and various target genes were acquired by quantitative RT-PCR using specific universal probe library primer probe sets (Roche) and a Roche Light Cycler II 480.

Article Title: Effect of the sonic hedgehog receptor smoothened on the survival and function of dopaminergic neurons
Article Snippet: Brain tissues were harvested for qRT-PCR analysis at 1-hour post last methamphetamine injection (D5) in wt and DAT-Smo ko mice (n=8 for each group). .. Total RNA (1 ug) was treated with RQ-1 Rnase-free Dnase I and reverse transcribed into cDNA using random hexamers by Superscript III reverse transcriptase (Life Sciences). cDNA levels for HPRT1 (hypoxanthine phosphoribosyltransferase 1), Hmbs (hydroxymethylbilane synthase) and various target genes were determined, using specific universal probe Library primer probe sets (Roche), by quantitative RT-PCR using a Roche Light Cycler II 480.

Injection:

Article Title: Dopaminergic neuron-specific deletion of p53 gene is neuroprotective in an experimental Parkinson's disease model
Article Snippet: Tissues were harvested for qRT-PCR analysis at 48 hours after the last MPTP or saline injection. .. Total RNA (0.7 μg) was treated with RQ-1 Rnase-free Dnase I and reverse transcribed into cDNA using random hexamers by Superscript III reverse transcriptase (Life Sciences). cDNA levels for HPRT1 (hypoxanthine phosphoribosyltransferase 1), Hmbs (hydroxymethylbilane synthase) and various target genes were determined using specific universal probe Library primer probe sets (Roche) by quantitative RT-PCR using a Roche Light Cycler II 480.

Article Title: Effect of the sonic hedgehog receptor smoothened on the survival and function of dopaminergic neurons
Article Snippet: Brain tissues were harvested for qRT-PCR analysis at 1-hour post last methamphetamine injection (D5) in wt and DAT-Smo ko mice (n=8 for each group). .. Total RNA (1 ug) was treated with RQ-1 Rnase-free Dnase I and reverse transcribed into cDNA using random hexamers by Superscript III reverse transcriptase (Life Sciences). cDNA levels for HPRT1 (hypoxanthine phosphoribosyltransferase 1), Hmbs (hydroxymethylbilane synthase) and various target genes were determined, using specific universal probe Library primer probe sets (Roche), by quantitative RT-PCR using a Roche Light Cycler II 480.

Software:

Article Title: A Rho-actin signaling pathway shapes cell wall boundaries in Arabidopsis xylem vessels
Article Snippet: After reverse transcription with oligo(dT)20 priming and SuperScript III reverse transcriptase, quantitative reverse transcription PCR was performed using a LightCycler 96 Instrument (Roche Diagnostics) with FastStart Essential DNA Green Master (Roche Diagnostics). .. Quantification of gene expression was performed using LightCycler software, and expression levels were determined relative to UBQ10 .

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