The severe acute respiratory syndrome (SARS) has recently been identified as a new clinical entit... more The severe acute respiratory syndrome (SARS) has recently been identified as a new clinical entity. SARS is thought to be caused by an unknown infectious agent.
We have developed a novel "real time" quantitative PCR method. The method measures PCR product ac... more We have developed a novel "real time" quantitative PCR method. The method measures PCR product accumulation through a dual-labeled fluorogenic probe {i.e., TaqMan Probe}. This method provides very accurate and reproducible quantitation of gene copies. Unlike other quantitative PCR methods, real-time PCR does not require post-PCR sample handling, preventing potential PCR product carry-over contamination and resulting in much faster and higher throughput assays. The real-time PCR method has a very large dynamic range of starting target molecule determination (at least five orders of magnitude}. Real-time quantitative PCR is extremely accurate and less labor-intensive than current quantitative PCR methods.
A novel approach to quantitative reverse transcriptase polymerase chain reaction (QC RT-PCR} usin... more A novel approach to quantitative reverse transcriptase polymerase chain reaction (QC RT-PCR} using real time detection and the S' nuclease assay has been developed. Cystic fibrosis transmembrane transductance regulator (CFTR} target mRNA is reverse transcribed, amplified, detected, and quantitated in real time. A fluorogenic probe was designed to detect the CFTR amplicon. Relative increase in 6-carboxy-fluorescein reporter fluorescent emission is monitored during PCR amplification using an analytical thermal cycler. An internal control template containing the same primer sequences as the CFTR amplicon, but a different internal sequence, has been designed as a control. An internal control probe with a reporter fluorescent dye tetrachloro-6-carboxy-fluorescein was designed to hybridize to the internal control amplicon. The internal control template is placed in each reaction tube and is used for quantitative analysis of the CFTR mRNA. This method provides a convenient and high-throughput format for QC RT-PCR.
The severe acute respiratory syndrome (SARS) has recently been identified as a new clinical entit... more The severe acute respiratory syndrome (SARS) has recently been identified as a new clinical entity. SARS is thought to be caused by an unknown infectious agent.
We have developed a novel "real time" quantitative PCR method. The method measures PCR product ac... more We have developed a novel "real time" quantitative PCR method. The method measures PCR product accumulation through a dual-labeled fluorogenic probe {i.e., TaqMan Probe}. This method provides very accurate and reproducible quantitation of gene copies. Unlike other quantitative PCR methods, real-time PCR does not require post-PCR sample handling, preventing potential PCR product carry-over contamination and resulting in much faster and higher throughput assays. The real-time PCR method has a very large dynamic range of starting target molecule determination (at least five orders of magnitude}. Real-time quantitative PCR is extremely accurate and less labor-intensive than current quantitative PCR methods.
A novel approach to quantitative reverse transcriptase polymerase chain reaction (QC RT-PCR} usin... more A novel approach to quantitative reverse transcriptase polymerase chain reaction (QC RT-PCR} using real time detection and the S' nuclease assay has been developed. Cystic fibrosis transmembrane transductance regulator (CFTR} target mRNA is reverse transcribed, amplified, detected, and quantitated in real time. A fluorogenic probe was designed to detect the CFTR amplicon. Relative increase in 6-carboxy-fluorescein reporter fluorescent emission is monitored during PCR amplification using an analytical thermal cycler. An internal control template containing the same primer sequences as the CFTR amplicon, but a different internal sequence, has been designed as a control. An internal control probe with a reporter fluorescent dye tetrachloro-6-carboxy-fluorescein was designed to hybridize to the internal control amplicon. The internal control template is placed in each reaction tube and is used for quantitative analysis of the CFTR mRNA. This method provides a convenient and high-throughput format for QC RT-PCR.
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