RNA Gene Expression For Multiplex

Combining Luminex xMAP technology with branched DNA technology, i-DNA offers RNA quantitation of up to 36 different targets per sample, per well of 96-well plate using QuantiGene Plex 2.0 (QGP) Assay.

QGP Assay is a hybridization-based assay using the xMAP® Luminex® magnetic beads and performed on 96-well plates. The assay is based on DIRECT quantification of the RNA targets using xMAP Luminex beads for multiplexing of 3 to 36 RNA targets and branched DNA (bDNA) signal amplification technology. On the first day the sample is lysed to release the RNAs, and incubated overnight with target specific probe sets panel. On the second day the signal amplification tree is build via sequential hybridization of PreAmplifier, Amplifier and Label Probe. Each amplification unit gives a 400 x signal amplification and there are six amplification units per target RNA copy leaning to a 2,400 x signal amplification per copy RNA. The signal is detected by adding SAPE substrate and using a Luminex instrument for the read out.

Assay specifications:

Limit of Detection:
≤ 1,000 – 2,000 transcripts/assay well

Limit of Quantitation:
≤ 2,000 – 4,000 transcripts/assay well

Linear Dynamic Range:
≥ 3.0 logs

Assay CV:
≤ 15% intra-assay; ≤ 20% inter-assay

Compatible Sample Types:
Cultured cells, bacteria, whole blood, PAXgene blood or dried blood spots, fresh/frozen tissues (animal or plant), FFPE samples, purified RNA

Assay Format:
96-well plate

Targets/well:
3 – 36

Key benefits are:

Sensitivity comparable to real time PCR, limit of detection ≤ 1,000 transcripts/assay well
RNA quantitation directly from cultured cell or whole blood lysates or fresh, frozen or formalin-fixed, paraffin-embedded (FFPE) tissue homogenates
No RNA purification
No reverse transcription
No target amplification
Simple assay workflow, significantly reduce turnaround time
Widely used in biomarker discovery, secondary screening, microarray validation, quantification of RNAi knockdowns and predictive toxicology

i-DNA offers RNA gene expression service starting from any of these sample types – Cultured Cells, whole blood, PAXgene blood or dried blood spots, fresh/frozen tissues, FFPE samples, purified RNA.

STEP

Customer provides the samples and GenBank accession number of target gene(s).Up to 36 different genes can be multiplexed into one panel.

STEP

i-DNA designs specific probe set against the target gene and synthesize it.

STEP

i-DNA performs the RNA quantitation and provides customer the date/report.

Note: QuantiGene Plex bDNA-based gene expression solution demonstrates clear performance benefits over PCR based technologies in an independent FDA MAQC validation study published in Nature Biotechnology 2006, Vol. 24, p.1115-22. Subsequently, there are numerous papers published using QuantiGene Plex. Contact i-DNA for the up-to-date publication list.

Consider outsourcing your RNA gene expression project, please contact i-DNA for a no-obligation consultation. All information provided will be kept strictly confidential.