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Bio-Rad cdna fragment levels
Comparison different PCR methods and impact on quantification. Graphic representation of the quantification of dystrophin <t>cDNA</t> construct templates lacking or containing exon 51 mixed in different ratios ranging from 0% to 100% using nested PCR (40, 50 and 60 cycles), <t>qPCR</t> (LinRegPCR and Pfaffl methods) and ddPCR. The dashed red line indicates the theoretical template ratios. The solid black ddPCR line is almost indistinguishable from the theoretical line and both lines are superimposed. Data is based on one experiment using 2 replicates for ddPCR and 3 replicates for qPCR.
Cdna Fragment Levels, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Digital Droplet PCR for the Absolute Quantification of Exon Skipping Induced by Antisense Oligonucleotides in (Pre-)Clinical Development for Duchenne Muscular Dystrophy"

Article Title: Digital Droplet PCR for the Absolute Quantification of Exon Skipping Induced by Antisense Oligonucleotides in (Pre-)Clinical Development for Duchenne Muscular Dystrophy

Journal: PLoS ONE

doi: 10.1371/journal.pone.0162467

Comparison different PCR methods and impact on quantification. Graphic representation of the quantification of dystrophin cDNA construct templates lacking or containing exon 51 mixed in different ratios ranging from 0% to 100% using nested PCR (40, 50 and 60 cycles), qPCR (LinRegPCR and Pfaffl methods) and ddPCR. The dashed red line indicates the theoretical template ratios. The solid black ddPCR line is almost indistinguishable from the theoretical line and both lines are superimposed. Data is based on one experiment using 2 replicates for ddPCR and 3 replicates for qPCR.
Figure Legend Snippet: Comparison different PCR methods and impact on quantification. Graphic representation of the quantification of dystrophin cDNA construct templates lacking or containing exon 51 mixed in different ratios ranging from 0% to 100% using nested PCR (40, 50 and 60 cycles), qPCR (LinRegPCR and Pfaffl methods) and ddPCR. The dashed red line indicates the theoretical template ratios. The solid black ddPCR line is almost indistinguishable from the theoretical line and both lines are superimposed. Data is based on one experiment using 2 replicates for ddPCR and 3 replicates for qPCR.

Techniques Used: Polymerase Chain Reaction, Construct, Nested PCR, Real-time Polymerase Chain Reaction

Related Articles

Real-time Polymerase Chain Reaction:

Article Title: Digital Droplet PCR for the Absolute Quantification of Exon Skipping Induced by Antisense Oligonucleotides in (Pre-)Clinical Development for Duchenne Muscular Dystrophy
Article Snippet: .. In each qPCR run, a no template control (NTC) was included as negative control. cDNA fragment levels were determined on a CFX96 real-time thermal cycler (BioRad) using the following protocol: 95°C for 10 min, then 40 cycles of 95°C for 15 sec and 60°C for 1 min. qPCR data was analyzed using both the Pfaffl method [ ] and the LinRegPCR method [ ]. .. The Pfaffl calculation was performed using the efficiencies derived from the standard curves for both assays (with and without exon 51 skip) in the following manner: ratio of skipped versus non-skipped = E (skipped)-Cq (skipped) /E (non-skipped)-Cq (non-skipped) .

Negative Control:

Article Title: Digital Droplet PCR for the Absolute Quantification of Exon Skipping Induced by Antisense Oligonucleotides in (Pre-)Clinical Development for Duchenne Muscular Dystrophy
Article Snippet: .. In each qPCR run, a no template control (NTC) was included as negative control. cDNA fragment levels were determined on a CFX96 real-time thermal cycler (BioRad) using the following protocol: 95°C for 10 min, then 40 cycles of 95°C for 15 sec and 60°C for 1 min. qPCR data was analyzed using both the Pfaffl method [ ] and the LinRegPCR method [ ]. .. The Pfaffl calculation was performed using the efficiencies derived from the standard curves for both assays (with and without exon 51 skip) in the following manner: ratio of skipped versus non-skipped = E (skipped)-Cq (skipped) /E (non-skipped)-Cq (non-skipped) .

Size-exclusion Chromatography:

Article Title: Digital Droplet PCR for the Absolute Quantification of Exon Skipping Induced by Antisense Oligonucleotides in (Pre-)Clinical Development for Duchenne Muscular Dystrophy
Article Snippet: .. In each qPCR run, a no template control (NTC) was included as negative control. cDNA fragment levels were determined on a CFX96 real-time thermal cycler (BioRad) using the following protocol: 95°C for 10 min, then 40 cycles of 95°C for 15 sec and 60°C for 1 min. qPCR data was analyzed using both the Pfaffl method [ ] and the LinRegPCR method [ ]. .. The Pfaffl calculation was performed using the efficiencies derived from the standard curves for both assays (with and without exon 51 skip) in the following manner: ratio of skipped versus non-skipped = E (skipped)-Cq (skipped) /E (non-skipped)-Cq (non-skipped) .

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    Bio-Rad cdna fragment levels
    Comparison different PCR methods and impact on quantification. Graphic representation of the quantification of dystrophin <t>cDNA</t> construct templates lacking or containing exon 51 mixed in different ratios ranging from 0% to 100% using nested PCR (40, 50 and 60 cycles), <t>qPCR</t> (LinRegPCR and Pfaffl methods) and ddPCR. The dashed red line indicates the theoretical template ratios. The solid black ddPCR line is almost indistinguishable from the theoretical line and both lines are superimposed. Data is based on one experiment using 2 replicates for ddPCR and 3 replicates for qPCR.
    Cdna Fragment Levels, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cdna fragment levels/product/Bio-Rad
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cdna fragment levels - by Bioz Stars, 2020-07
    90/100 stars
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    Comparison different PCR methods and impact on quantification. Graphic representation of the quantification of dystrophin cDNA construct templates lacking or containing exon 51 mixed in different ratios ranging from 0% to 100% using nested PCR (40, 50 and 60 cycles), qPCR (LinRegPCR and Pfaffl methods) and ddPCR. The dashed red line indicates the theoretical template ratios. The solid black ddPCR line is almost indistinguishable from the theoretical line and both lines are superimposed. Data is based on one experiment using 2 replicates for ddPCR and 3 replicates for qPCR.

    Journal: PLoS ONE

    Article Title: Digital Droplet PCR for the Absolute Quantification of Exon Skipping Induced by Antisense Oligonucleotides in (Pre-)Clinical Development for Duchenne Muscular Dystrophy

    doi: 10.1371/journal.pone.0162467

    Figure Lengend Snippet: Comparison different PCR methods and impact on quantification. Graphic representation of the quantification of dystrophin cDNA construct templates lacking or containing exon 51 mixed in different ratios ranging from 0% to 100% using nested PCR (40, 50 and 60 cycles), qPCR (LinRegPCR and Pfaffl methods) and ddPCR. The dashed red line indicates the theoretical template ratios. The solid black ddPCR line is almost indistinguishable from the theoretical line and both lines are superimposed. Data is based on one experiment using 2 replicates for ddPCR and 3 replicates for qPCR.

    Article Snippet: In each qPCR run, a no template control (NTC) was included as negative control. cDNA fragment levels were determined on a CFX96 real-time thermal cycler (BioRad) using the following protocol: 95°C for 10 min, then 40 cycles of 95°C for 15 sec and 60°C for 1 min. qPCR data was analyzed using both the Pfaffl method [ ] and the LinRegPCR method [ ].

    Techniques: Polymerase Chain Reaction, Construct, Nested PCR, Real-time Polymerase Chain Reaction