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This can be challenging when working with some biological samples.
Fortunately, a number of commercial products have been developed to facilitate the isolation of nucleic acids in high purity.
Because it takes several cycles for enough product to be readily detectable, the plot of fluorescence vs. At later cycles, the reaction substrates become depleted, PCR product no longer doubles, and the curve begins to flatten.
However, its limitations are emphasised by the considerable prognostic heterogeneity of patients within a given tumour stage: not all patients with LN-negative cancers are cured and not all patients with LN-positive tumours die from their disease.
Library of targeted real time PCR primer sets Perform up to 200 PCR Arrays! (Based on a 10 ul reaction volume)Flexibility to design one's own experiments Microplate containing 88 targeted plus 8 housekeeping gene primer sets (20ul per well, 10u M concentration)Note: Primer sequences unavailable for primer libraries If you would like to request a specific library for a pathway or organism not listed, please let us know: [email protected] goal is to provide a cost-effective approach to validate microarray data and quantitate gene expression using real time PCR assays Perform up to 1000 assays Detailed information on optimal reaction conditions provided Primer sequences provided!
Overview - Quantitative "Real Time" PCRQuantitative real time PCR has revolutionized our ability to measure nucleic acid concentrations and is an important step for the validation of expression data generated by microarray analysis and other genomics techniques.
This has resulted in a search for more accurate staging protocols and has seen the introduction of the concept of "molecular staging", the incorporation of molecular parameters into clinical tumour staging.
Quantification of disease-associated m RNA is one such parameter that utilises the q RT-PCR assay's potential for generating quantitative results.
The plot of Ct versus template is linear, thus a comparison of Ct values between multiple reactions enables one to calculate the concentration of the target nucleic acid.