Affymetrix | Panomics Solutions
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QuantiGene 2.0 DNA Assay: Overview

QuantiGene 2.0 DNA Assay - Benefits

  • Detect original DNA population - using signal amplification
    • No DNA purification
    • No reverse transcription
    • No PCR amplification
  • Avoid biases and sample loss - with direct from lysate quantitation using signal amplification
    • No target loss during DNA isolation
    • No biases associated with enzyme inhibition or fluorescent dye incorporation during PCR amplification
  • Simplified workflow - with direct from lysate quantitation
    • Lyse and go sample preparation
    • ELISA like protocol - add and wash reagetns
  • Sensitive quantitation
    • Sensitivity comparable to real time PCR (limit of detection)
    • Precisely quantify as littel as 20% difference in copy number
  • Analyze any sample, any target
    • DNA copy number quantitation directly from any sample - cultured cells; whole blood; dried blood spots; fresh, frozen or formalin-fixed, paraffin-embedded (FFPE) tissues; plants; bacteria; purified DNA. 
    • Create probe sets against any target sequence
  • Read using a standard plate luminometer


QuantiGene 2.0 - Applications

  • DNA Assay - used for copy number determination in transgenic animals, plant zygocity testing.


QuantiGene 2.0 Assay Workflow

QuantiGene assays bring the power of branched DNA (bDNA) technology to basic and pre-clinical research. The bDNA assay is a sandwich nucleic acid hybridization method that uses bDNA molecules to amplify signal from captured target RNA. bDNA technology forms the basis of the FDA-approved clinical diagnostic VERSANT 3.0 assays for HIV, HCV and HBV viral load, that are offered commercially by Siemens and have been in use for over a decade. Another advantage of bDNA assays is that RNA is measured directly from the sample source, without RNA purification or enzymatic manipulation, thereby avoiding inefficiencies and variability introduced by or errors inherent to these processes.

Assay Specifications

  Limit of Detection   ≤ 10,000 copies/assay wella
  Limit of Quantitation   ≤ 20,000 copies/assay wellb
  Linear Dynamic Range   ≥ 2.5 logsc
  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(human, animal or plant),
  FFPE samples, purified DNA
  Assay Format   96-well plated
  Targets/well   1 target per well
  Automation Compatible
  Yese

a Defined as signal greater than background plus three standard deviations of the background.
b Defined as the signal just above the lowest signal that obtains an 80-120% spike recovery.
c Defined as the assay window that consistently achieves a 80-120% accuracy of fold change.
d Contact Affymetrix for information on a 384-well version.
e Batch or full automation using standard automation equipment. Contact Technical Support for protocol details and assistance in setting up your automation system.