superscript iii rt pcr platinum taq high fidelity kit  (Thermo Fisher)


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    Name:
    SuperScript III One Step RT PCR System
    Description:
    The SuperScript III One Step RT PCR System with Platinum Taq DNA Polymerase is designed for the sensitive reproducible end point detection and analysis of RNA molecules by RT PCR Using this convenient one step formulation you can perform both cDNA synthesis and PCR amplification in a single tube using gene specific primers and target RNAs from either total RNA or mRNA The system uses a mixture of SuperScript III Reverse Transcriptase and Platinum Taq DNA polymerase in an optimized reaction buffer and it can detect a wide range of RNA targets from 200 bp to 4 5 kb The amount of starting material can range from 0 01 pg to 1 µg of total RNA Key advantages of this kit • Sensitive routine detection down to 0 01 pg total RNA see figure • Convenience one step format for speed convenience and less reaction to reaction variability• Specificity uses SuperScript III RT for cDNA synthesis up to 55°C for more specific priming with gene specific primers see figure • Amplicon size compatible detection of targets up to 4 5 kb in length for greater flexibilitySuperScript III Reverse TranscriptaseSuperScript III Reverse Transcriptase is a version of M MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability The enzyme can synthesize cDNA at a temperature range of 45 60°C providing increased specificity higher yields of cDNA and more full length product than other reverse transcriptases Because SuperScript III RT is not significantly inhibited by ribosomal and transfer RNA it can be used to synthesize cDNA from total RNA Platinum Taq DNA polymerasePlatinum Taq DNA polymerase is recombinant Taq DNA polymerase complexed with a proprietary antibody that blocks polymerase activity at ambient temperatures Activity is restored after the denaturation step in PCR cycling at 94°C providing an automatic hot start in PCR for increased sensitivity specificity and yield
    Catalog Number:
    12574018
    Price:
    None
    Applications:
    One-Step RT-PCR|PCR & Real-Time PCR|RT-PCR|Reverse Transcription
    Category:
    Kits and Assays
    Buy from Supplier


    Structured Review

    Thermo Fisher superscript iii rt pcr platinum taq high fidelity kit
    ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all <t>three</t> of the cell populations were subjected to <t>RT-PCR</t> to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).
    The SuperScript III One Step RT PCR System with Platinum Taq DNA Polymerase is designed for the sensitive reproducible end point detection and analysis of RNA molecules by RT PCR Using this convenient one step formulation you can perform both cDNA synthesis and PCR amplification in a single tube using gene specific primers and target RNAs from either total RNA or mRNA The system uses a mixture of SuperScript III Reverse Transcriptase and Platinum Taq DNA polymerase in an optimized reaction buffer and it can detect a wide range of RNA targets from 200 bp to 4 5 kb The amount of starting material can range from 0 01 pg to 1 µg of total RNA Key advantages of this kit • Sensitive routine detection down to 0 01 pg total RNA see figure • Convenience one step format for speed convenience and less reaction to reaction variability• Specificity uses SuperScript III RT for cDNA synthesis up to 55°C for more specific priming with gene specific primers see figure • Amplicon size compatible detection of targets up to 4 5 kb in length for greater flexibilitySuperScript III Reverse TranscriptaseSuperScript III Reverse Transcriptase is a version of M MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability The enzyme can synthesize cDNA at a temperature range of 45 60°C providing increased specificity higher yields of cDNA and more full length product than other reverse transcriptases Because SuperScript III RT is not significantly inhibited by ribosomal and transfer RNA it can be used to synthesize cDNA from total RNA Platinum Taq DNA polymerasePlatinum Taq DNA polymerase is recombinant Taq DNA polymerase complexed with a proprietary antibody that blocks polymerase activity at ambient temperatures Activity is restored after the denaturation step in PCR cycling at 94°C providing an automatic hot start in PCR for increased sensitivity specificity and yield
    https://www.bioz.com/result/superscript iii rt pcr platinum taq high fidelity kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    superscript iii rt pcr platinum taq high fidelity kit - by Bioz Stars, 2020-09
    99/100 stars

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    Images

    1) Product Images from "Transposon Mutagenesis of the Zika Virus Genome Highlights Regions Essential for RNA Replication and Restricted for Immune Evasion"

    Article Title: Transposon Mutagenesis of the Zika Virus Genome Highlights Regions Essential for RNA Replication and Restricted for Immune Evasion

    Journal: Journal of Virology

    doi: 10.1128/JVI.00698-17

    ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all three of the cell populations were subjected to RT-PCR to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).
    Figure Legend Snippet: ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all three of the cell populations were subjected to RT-PCR to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).

    Techniques Used: Mutagenesis, Library Screening, Sequencing, Transfection, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Infection

    Related Articles

    Amplification:

    Article Title: In vitro and in vivo anti-tumor activity of CoQ0 against melanoma cells: inhibition of metastasis and induction of cell-cycle arrest and apoptosis through modulation of Wnt/β-catenin signaling pathways
    Article Snippet: .. A 1 μg sample of total RNA was subjected to RT-PCR using a BioRad iCycler PCR instrument (Bio-Rad, Hercules, CA) and the SuperScript-III® One-Step RT-PCR Platinum taq ® Kit (Invitrogen); amplification was performed in 30-38 cycles at 94°C for 45s (denaturing), 60-65°C for 45s (annealing), and 72°C for 1 min (primer extension). ..

    Article Title: Taenia solium glutathione transferase fraction activates macrophages and favors the development of Th1-type response
    Article Snippet: .. For reverse-transcription PCR (RT-PCR), 1 μg of total RNA was retro-transcribed and amplified with a couple of gen-specific primers at 50 pM using the SuperScript III One-Step RT-PCR System kit (Invitrogen). .. Glyceraldehyde-3-phosphate dehydrogenase (Gapdh ) (housekeeping gene), Arg1 , nitric oxide synthase 2 inducible (Nos2 ), and Chil3 genes were amplified using the primers designed previously [ ].

    Synthesized:

    Article Title: An Xist-activating antisense RNA required for X-chromosome inactivation
    Article Snippet: .. Amplicons containing SNPs were designed using the PyroMark Assay Design software. cDNAs were synthesized using Invitrogen SuperScript III One-Step RT–PCR System (Invitrogen, #12574-026). .. Following the PCR reaction, 5 μl of a total of 25 μl was run on a 3% agarose gel to assess the efficacy of the RT and amplification.

    Quantitative RT-PCR:

    Article Title: Divergent Simian Arteriviruses Cause Simian Hemorrhagic Fever of Differing Severities in Macaques
    Article Snippet: .. Briefly, viral RNA was reverse transcribed and quantified using the SuperScript III one-step qRT-PCR system (Invitrogen, Carlsbad, CA) on a LightCycler 480 (Roche, Indianapolis, IN). .. The reaction mixture contained MgSO4 at a final concentration of 3.0 mM, 150 ng of random primers (Promega), amplification primers at a concentration of 600 nM (5′-ACACGGCTACCCTTACTCC-3′ and 5′-TCGAGGTTAARCGGTTGAGA-3′), and probe (5′-Quasar-670-TTCTGGTCCTCTTGCGAAGGC-black hole quencher 2 [BHQ2]-3′) at a concentration of 100 nM.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: In vitro and in vivo anti-tumor activity of CoQ0 against melanoma cells: inhibition of metastasis and induction of cell-cycle arrest and apoptosis through modulation of Wnt/β-catenin signaling pathways
    Article Snippet: .. A 1 μg sample of total RNA was subjected to RT-PCR using a BioRad iCycler PCR instrument (Bio-Rad, Hercules, CA) and the SuperScript-III® One-Step RT-PCR Platinum taq ® Kit (Invitrogen); amplification was performed in 30-38 cycles at 94°C for 45s (denaturing), 60-65°C for 45s (annealing), and 72°C for 1 min (primer extension). ..

    Article Title: AKAP13 Rho-GEF and PKD-Binding Domain Deficient Mice Develop Normally but Have an Abnormal Response to ?-Adrenergic-Induced Cardiac Hypertrophy
    Article Snippet: .. Total RNA was extracted from ES cells with Trizol (Invitrogen), and RT-PCR was conducted using the SuperScript III One-Step RT-PCR kit (Invitrogen). .. Forward primers for RT-PCR were designed using Primer3 ( ) .

    Article Title: Metastasis-associated phosphatase PRL-2 regulates tumor cell migration and invasion
    Article Snippet: .. Reverse transcription-PCR (RT-PCR) for PRL-1, PRL-2, PRL-3, and actin as an internal control was carried out in a volume of 50 µL by SuperScript III One-step RT-PCR System (Invitrogen) as per manufacturer's instruction. ..

    Article Title: Taenia solium glutathione transferase fraction activates macrophages and favors the development of Th1-type response
    Article Snippet: .. For reverse-transcription PCR (RT-PCR), 1 μg of total RNA was retro-transcribed and amplified with a couple of gen-specific primers at 50 pM using the SuperScript III One-Step RT-PCR System kit (Invitrogen). .. Glyceraldehyde-3-phosphate dehydrogenase (Gapdh ) (housekeeping gene), Arg1 , nitric oxide synthase 2 inducible (Nos2 ), and Chil3 genes were amplified using the primers designed previously [ ].

    Article Title: Non-canonical translation initiation of the spliced mRNA encoding the human T-cell leukemia virus type 1 basic leucine zipper protein
    Article Snippet: .. The RT-PCR assay was carried out using the SuperScript™ III One-Step RT-PCR System with Platinum® Taq DNA polymerase kit (#52122; ThermoFisher Scientific) according to manufacturer’s protocol, 5 ng of total RNA, and the primers Pforluc and p2anti as described in ( , ). ..

    Article Title: An Xist-activating antisense RNA required for X-chromosome inactivation
    Article Snippet: .. Amplicons containing SNPs were designed using the PyroMark Assay Design software. cDNAs were synthesized using Invitrogen SuperScript III One-Step RT–PCR System (Invitrogen, #12574-026). .. Following the PCR reaction, 5 μl of a total of 25 μl was run on a 3% agarose gel to assess the efficacy of the RT and amplification.

    Article Title: Functional and Structural Analysis of the Internal Ribosome Entry Site Present in the mRNA of Natural Variants of the HIV-1
    Article Snippet: .. The RT-PCR assay was carried out with SuperScript™ III One-Step RT-PCR System using Platinum® Taq DNA polymerase (Invitrogen) according to the manufacturer’s protocol, using 1 µg of RNA and the primers described above. .. The RT-qPCR assay was conducted using the primers RLucS ( 5 ′ -AGGTGAAGTTCGTCGTCCAACATTATC-3 ′ ) and RLucAS ( 5 ′ -GAAACTTCTTGGCACCTTCAACAATAGC-3 ′ ) for RLuc amplification (193bp); or FLucS ( 5 ′ - ACTTCGAAATGTCCGTTCGG-3 ′ ) and FLucAS ( 5 ′ - GCAACTCCGATAAATAACGCG-3 ′ ) for FLuc amplification (135bp), using the Brilliant® II SYBR® Green QRT-PCR Master Mix KIT (Stratagene, Agilent Technologies Company, Santa Clara, CA, USA).

    Polymerase Chain Reaction:

    Article Title: In vitro and in vivo anti-tumor activity of CoQ0 against melanoma cells: inhibition of metastasis and induction of cell-cycle arrest and apoptosis through modulation of Wnt/β-catenin signaling pathways
    Article Snippet: .. A 1 μg sample of total RNA was subjected to RT-PCR using a BioRad iCycler PCR instrument (Bio-Rad, Hercules, CA) and the SuperScript-III® One-Step RT-PCR Platinum taq ® Kit (Invitrogen); amplification was performed in 30-38 cycles at 94°C for 45s (denaturing), 60-65°C for 45s (annealing), and 72°C for 1 min (primer extension). ..

    Article Title: Taenia solium glutathione transferase fraction activates macrophages and favors the development of Th1-type response
    Article Snippet: .. For reverse-transcription PCR (RT-PCR), 1 μg of total RNA was retro-transcribed and amplified with a couple of gen-specific primers at 50 pM using the SuperScript III One-Step RT-PCR System kit (Invitrogen). .. Glyceraldehyde-3-phosphate dehydrogenase (Gapdh ) (housekeeping gene), Arg1 , nitric oxide synthase 2 inducible (Nos2 ), and Chil3 genes were amplified using the primers designed previously [ ].

    Pyromark Assay:

    Article Title: An Xist-activating antisense RNA required for X-chromosome inactivation
    Article Snippet: .. Amplicons containing SNPs were designed using the PyroMark Assay Design software. cDNAs were synthesized using Invitrogen SuperScript III One-Step RT–PCR System (Invitrogen, #12574-026). .. Following the PCR reaction, 5 μl of a total of 25 μl was run on a 3% agarose gel to assess the efficacy of the RT and amplification.

    Software:

    Article Title: An Xist-activating antisense RNA required for X-chromosome inactivation
    Article Snippet: .. Amplicons containing SNPs were designed using the PyroMark Assay Design software. cDNAs were synthesized using Invitrogen SuperScript III One-Step RT–PCR System (Invitrogen, #12574-026). .. Following the PCR reaction, 5 μl of a total of 25 μl was run on a 3% agarose gel to assess the efficacy of the RT and amplification.

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  • 99
    Thermo Fisher superscript iii rt pcr platinum taq high fidelity kit
    ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all <t>three</t> of the cell populations were subjected to <t>RT-PCR</t> to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).
    Superscript Iii Rt Pcr Platinum Taq High Fidelity Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript iii rt pcr platinum taq high fidelity kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    superscript iii rt pcr platinum taq high fidelity kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

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    ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all three of the cell populations were subjected to RT-PCR to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).

    Journal: Journal of Virology

    Article Title: Transposon Mutagenesis of the Zika Virus Genome Highlights Regions Essential for RNA Replication and Restricted for Immune Evasion

    doi: 10.1128/JVI.00698-17

    Figure Lengend Snippet: ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all three of the cell populations were subjected to RT-PCR to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).

    Article Snippet: The viral RNA was then amplified in three segments by utilizing a SuperScript III RT-PCR Platinum Taq High Fidelity kit (Thermo Fisher Scientific).

    Techniques: Mutagenesis, Library Screening, Sequencing, Transfection, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Infection