ExProfile™ Pathway Gene qPCR Arrays

How qPCR Arrays Work

Figure 1. Gene qPCR array experimental work flow.

Key advantages

Close pathway correlation

• Genes are carefully chosen for their close pathway correlation based on a thorough literature search of peer-reviewed publications

Robust performance

• 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

Validated primers

• Each primer is designed using a proprietary algorithm and has been experimentally validated

Custom qPCR Arrays: 

GeneCopoeia offers 96-well and 384-well custom array types and each array type has 6 layouts to choose from depending on the number of genes and replicates to be analyzed. Different controls can also be included in the arrays to help monitor the sample quality, RT and PCR reaction efficiencies and replicates reproducibility.

• Genomic DNA control (GDC): detects genomic DNA contamination.
• Spike-in reverse transcription control (RT): monitors the efficiency of the RT reaction.
• Positive PCR control (PCR): verifies the PCR efficiency.
• Housekeeping genes (HK): can be used as endogenous positive controls and for array normalization.

For the best performance, All-in-One™ First Strand cDNA Synthesis Kit and All-in-One™ qPCR Mix are the recommended and supported reagents for use with these arrays.

All-in-One qPCR validated primers 

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.

Amplification curves
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).

Related arrays

• ExProfile™ cancer gene qPCR Arrays
• ExProfile™ disease and gene group qPCR Arrays
• ExProfile™ custom gene qPCR Array
• miProfile^TM miRNA qPCR arrays

Related reagents

• RNAzol®RT RNA isolation reagent
• All-in-One™ First-Strand cDNA Synthesis Kits
• All-in-One™ qPCR mix

ExProfile™ gene qPCR array user manual

All-in-One™ first-strand cDNA synthesis kit user manual

RNAzol® RT RNA isolation reagent user manual