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


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    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, 2020-05
    99/100 stars

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    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: TNFα blockade prevents the development of inflammatory bowel disease in HLA-B27 transgenic rats
    Article Snippet: .. One microgram of total RNA was reverse transcribed and amplified using the SuperScrip III One-Step RT-PCR System with Platinum Taq DNA Polymerase (Invitrogen). ..

    Article Title: Galectin-1 Regulates Tissue Exit of Specific Dendritic Cell Populations *
    Article Snippet: .. Real Time Quantitative RT-PCR cDNA was prepared from human DCs using the RNeasy mini kit (Qiagen) and the Super-Script III One-Step RT-PCR kit (Invitrogen). cDNA was amplified with specific primer sets using the LightCycler® 480 SYBR Green I Master reagent (Roche Applied Science) with the LightCycler® 480 (Roche Applied Science) and its detection software. ..

    Quantitative RT-PCR:

    Article Title: Galectin-1 Regulates Tissue Exit of Specific Dendritic Cell Populations *
    Article Snippet: .. Real Time Quantitative RT-PCR cDNA was prepared from human DCs using the RNeasy mini kit (Qiagen) and the Super-Script III One-Step RT-PCR kit (Invitrogen). cDNA was amplified with specific primer sets using the LightCycler® 480 SYBR Green I Master reagent (Roche Applied Science) with the LightCycler® 480 (Roche Applied Science) and its detection software. ..

    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.

    Article Title: Subclinical Infection of Macaques and Baboons with A Baboon Simarterivirus
    Article Snippet: .. Reverse transcription and PCR were performed using the SuperScript III One-Step qRT-PCR system (Invitrogen, Carlsbad, CA, USA) on a LightCycler 480 (Roche, Indianapolis, IN, USA). .. The 50 µL-reaction mixture contained 5 µL of extracted RNA, MgSO4 at a final concentration of 3.0 mM, with the two amplification primers at a concentration of 500 nM and probe at a concentration of 100 nM.

    SYBR Green Assay:

    Article Title: Galectin-1 Regulates Tissue Exit of Specific Dendritic Cell Populations *
    Article Snippet: .. Real Time Quantitative RT-PCR cDNA was prepared from human DCs using the RNeasy mini kit (Qiagen) and the Super-Script III One-Step RT-PCR kit (Invitrogen). cDNA was amplified with specific primer sets using the LightCycler® 480 SYBR Green I Master reagent (Roche Applied Science) with the LightCycler® 480 (Roche Applied Science) and its detection software. ..

    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: TNFα blockade prevents the development of inflammatory bowel disease in HLA-B27 transgenic rats
    Article Snippet: .. One microgram of total RNA was reverse transcribed and amplified using the SuperScrip III One-Step RT-PCR System with Platinum Taq DNA Polymerase (Invitrogen). ..

    Article Title: Galectin-1 Regulates Tissue Exit of Specific Dendritic Cell Populations *
    Article Snippet: .. Real Time Quantitative RT-PCR cDNA was prepared from human DCs using the RNeasy mini kit (Qiagen) and the Super-Script III One-Step RT-PCR kit (Invitrogen). cDNA was amplified with specific primer sets using the LightCycler® 480 SYBR Green I Master reagent (Roche Applied Science) with the LightCycler® 480 (Roche Applied Science) and its detection software. ..

    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: 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: Subclinical Infection of Macaques and Baboons with A Baboon Simarterivirus
    Article Snippet: .. Reverse transcription and PCR were performed using the SuperScript III One-Step qRT-PCR system (Invitrogen, Carlsbad, CA, USA) on a LightCycler 480 (Roche, Indianapolis, IN, USA). .. The 50 µL-reaction mixture contained 5 µL of extracted RNA, MgSO4 at a final concentration of 3.0 mM, with the two amplification primers at a concentration of 500 nM and probe at a concentration of 100 nM.

    Software:

    Article Title: Galectin-1 Regulates Tissue Exit of Specific Dendritic Cell Populations *
    Article Snippet: .. Real Time Quantitative RT-PCR cDNA was prepared from human DCs using the RNeasy mini kit (Qiagen) and the Super-Script III One-Step RT-PCR kit (Invitrogen). cDNA was amplified with specific primer sets using the LightCycler® 480 SYBR Green I Master reagent (Roche Applied Science) with the LightCycler® 480 (Roche Applied Science) and its detection software. ..

    Similar Products

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  • About
  • News
  • Press Release
  • Team
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  • Partners
  • Contact
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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, 2020-05
    99/100 stars
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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