ampliPHOX® Colorimetric Detection

ampliPHOX Colorimetric Detection Technology

ampliPHOX® Colorimetric Microarray Detection is an inexpensive alternative to traditional fluorescence detection for both nucleic acid and protein applications when paired with target-specific microarrays (less than 2,500 spots). Rather than a fluorescent label, ampliPHOX utilizes a photoinitiator that, when activated by light, generates signal amplification through polymerization of an organic monomer. The detection technology is easy to use and can be completed within minutes. The ampliPHOX Reader is used to activate photopolymerization and to image the resulting pattern of visual spots. Once programmed for your array, the software can also interpret the image – providing the user with immediate results.

Customer Testimonial
“After evaluating the ampliPHOX colorimetric technology, my lab no longer uses expensive fluorescent dyes for detection of pathogens with DNA microarrays. The low cost of the ampliPHOX reagents in conjunction with the simplicity and rapidness of this colorimetric assay has allowed us to increase the number of bacterial isolates to be genotyped with DNA microarrays. We are now getting in a several hours the results that took us days to obtain with our previous assays.”

–Beatriz Quiñones, PhD, Research Molecular Biologist, USDA-ARS, Western Reg. Res. Ctr., Albany CA

ampliPHOX Highlights

  • Achieve equivalent sensitivity to traditional fluorescence microarray detection methods at 10x lower price
  • Proprietary detection technology that provides visual, colorimetric results within 20 minutes with imaging and analysis performed by the ampliVIEW software
  • A flexible platform that can be optimized to detect proteins and nucleic acids, even within the same array.
  • Up to 500 unique targets can be tested in triplicate in a single array
  • Instrumentation is light weight (less than 3 lbs) and compact (5.8”H x 5.5”W x 5.7”L), and cost effective (less than $5,000)
  • Intuitive ampliVIEW software enables automated interpretation of user-designed arrays
  • ampliPHOX reagents are inexpensive and easy-to-use

ampliPHOX is intended for research and investigational applications only.

ampliPHOX Specifications
Excitation Source LED
Weight 2 lbs 5 oz
Dimensions (in) 5.5 (W) x 5.7 (D) x 5.8 (H)
Power Requirements 12 VDC 1.25 A
Environment 65-80ºF (18-27ºC)
Warranty One year (materials and workmanship)

ampliPHOX Recommendations:

  • For best results, always run a Full System Optimization with each new lot of ampliPHY that you receive and run a System/Reagent Check when opening a new aliquot of ampliPHY if that lot has already been optimized.  For more information about the Optimization & Reagent Check, refer to the most recent version of the ampliPHOX Operation Manual.
  • Be sure to regularly clean out your wash bins and humidity chamber.
  • The ampliPHY reagents are oxygen sensitive so be sure to keep the vials closed when not in use to avoid excessive oxygen exposure.  If you see evidence of polymerization in the vial, do not use the reagent.
  • If there is evidence of salt precipitation in the ampliTAG Buffer, gently heat the buffer in a 35-39 C waterbath for 5-10 minutes until the salts are dissolved.
  • Avoid touching the spotted arrays with your hands or pipette tips as this could dislodge spots.
  • Make sure there are no air bubbles in the solutions applied to the arrays.  We’ve found that gently tapping the slide helps to get rid of them.
  • Always be sure to use the reagents within their expiration.

Pre-ampliPHOX Recommendations:

  • For nucleic acid microarrays, the target must be biotinylated in order for ampliPHOX to detect.  We’ve found the best ways to incorporate biotin is by using biotinylated NTP/dNTP or biotinylated primer(s) in a RT-PCR protocol.
  • Ideally, a single stranded target of approximately 100-300 bp should be used for your nucleic acid arrays.  Therefore, further processing of your target may be necessary for optimal performance.  For example, if you are using PCR product that is 1000 bp in length, enzymatically digest the product to create a ssDNA and heat treat to fragment.
  • For protein microarrays, a block is frequently necessary to avoid non-specific binding.  In-lieu of blocking each array individually, incorporate your optimized block buffer in the sample dilution and/or detection antibody dilution.
  •  A water wash is discouraged during the pre-ampliPHOX steps for protein microarrays.
  • Each assay is unique and optimization of the hybridization, binding, and wash steps as well as the buffers used is necessary to attain your best sensitivity and specificity.  We provide some starting points for your pre-ampliPHOX protocol here, but it’s up to you to fully optimize your microarray protocol.

web counter