For focused group profiling of pathway-related gene expression
ExProfile™ Pathway Gene qPCR Arrays profile the expression of pathway-related genes, which are carefully chosen for their close pathway correlation based on a thorough literature search of peer-reviewed publications. In each 96-well catalog array plate there are up to 84 pairs of All-in-One™ qPCR primers, which have been pre-validated and deposited in designated wells. In each plate there also are 12 wells of controls for monitoring the efficiency of the entire experimental process: from reverse transcription to qPCR reaction.
How qPCR Arrays Work
Figure 1. Gene qPCR array experimental work flow.
Close pathway correlation
- Genes are carefully chosen for their close pathway correlation based on a thorough literature search of peer-reviewed publications
- Sensitive – Detects as low as 4 copies of RNA using ExProfile qPCR array and recommended reagents/conditions
- Broad linearity – Simultaneously detects mRNAs at different expression levels
- Reproducible – High reproducibility (R2> 0.99) for inter-array and intra-array replicates
- Each primer is designed using a proprietary algorithm and has been experimentally validated
The All-in-One™ qPCR human- mouse- or rat-specific primers are designed by a proprietary algorithm and validated† for precision performance. Primer validation includes melting curve to ensure amplification of the correct target DNA (Figure 2). When used in combination with the All-in-One™ SYBR® Green qPCR Mix, the All-in-One™ primers deliver reliable and reproducible high performance in quantitative PCR assays.
|Melting curves||Agarose Gel Electrophoresis Validation Result|
Figure 2. Fourty-five pairs of gene-specific All-in-One™ qPCR primers were experimentally validated to yield a single dissociation curve peak and to generate a single amplification of the correct size for the targeted genes. A cDNA pool, containing reverse transcribed products of total RNA from 10 different human tissue (lung, liver, testicle, ovary, spleen, brain, placenta, pancreas, heart and mammary), was used as the qPCR validation template. qPCR was performed using 0.2 µM primer with 2X All-in-One qPCR Mix. Reactions were incubated for 10 min. at 95°C, followed by 40 cycles of 95°C for 10 sec.; 60°C, 20 sec. and 72°C, 15 sec. using Bio-Rad iQ5 Instrument. At the end of the last cycle, the temperature was increased from 72 to 95°C to produce a melting curve. Amplification curves, melting curves and agarose gel electrophoresis (validation result for positive in odd-lane and no template control (NTC) in even lane) shown for the 10 samples.
* Non-specific amplification and absence of primer-dimers are ensured when All-in-One validated PCR primers and PCR mix are used together.
† Primers validated automatically for human, mouse and rat. Inquire for validation of other species.
Data analysis tool
Analyze the qPCR result using GeneCopoeia's online Data Analysis System (free).
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