FAQ's for Luminex 100 System:

How does the Luminex work?

The Luminex system can analyze immunoassays, complex genetic analysis, and enzymatic assays in one format. The reactants, (antibodies, oligonucleotides, substrates, etc.) are attached to unique fluorescent microscopic beads (microspheres). The unique fluorescent emission for a given microsphere enables the Luminex to identify discrete measurements of multiple microsphere based reactions from a single sample (Luminex, Austin TX).

How is the Luminex used for SNP detection?

The use of microsphere based DNA polymerase assays utilizing single base extension (SBE) or allele specific primer extension (ASPE), allows users to multiplex many SNP markers in a single assay. Briefly, this is achieved by coupling oligonucleotides, labeled with a fluorescent reporter probe, that compliment allele codes of the SNP in query to multi colored microspheres. The coupled bead and reaction are passed though a single channel, by flow cytometry, where two lasers (green and red) excite both the bead and fluorescent probe. The red laser analyzes the color of the bead while the green laser analyzes the reporter color. Both readings are combined in the detection of multiple SNPs. (Luminex, Austin TX) For a general schematic of reaction and application see fig 1 from (Iannone et al. 2000, Cytometry. 39:131-140).

 Fig. 1

How will I learn to use the Luminex?

The IGMC will provide user training on the instrument and software, there is an optional on line training from Mirai Bio for in depth software analysis methods.

How much will it cost to use the Luminex?

Through a generous grant from the USDA, IGMC will be able to provide the equipment and services to center users at an affordable rate. The rate to use the instrument only is $10.00/hour.

What supplies/services will the IGMC provide?

The Center will provide the calibration beads and materials to support the Luminex system. The Center will perform all system maintenance and provide users with technical and troubleshooting advice. The Center will also maintain all user data on our server for backup and store large data files. The Center can also supply consumables for assays at cost; however users have no obligation to purchase them from the center.  IGMC has a few stocks of beads prepared with zip codes attached, please contact the center for more information.

Does the IGMC have protocols for SNP detection using the Luminex?
There are established SBE and ASPE methods (Single Base Extension & Allele Specific Primer Extension), contact the Center.

What size microplate can I use?

The Luminex at IGMC is calibrated for use with 96, conical well plates which can be purchased from the Center. 
(Fisher Brand part # 08-408-225.)

How long will it take to run an assay?

The actual microplate assay will take less than an hour. Preparing samples for assays will need to be done in advance and may be a multiple day process, which users can do in their own labs or in the Center. We have equipment that may aid in assembling reactions, including the MJ Research Tetrad® thermocycler and Hydra® Microdispenser, which center users can utilize.

How can I get started?

Contact IGMC for more information.

Do you have a list of references for the Luminex?

http://www.miraibio.com/pdf/products/liquidarrays/LuminexBibliography.pdf

These websites may be helpful:

Luminex, www.luminexcorp.com

Mirai Bio, www.miraibio.com

USDA (BARC)  http://bldg6.arsusda.gov/~pooley/soy/cregan/lum1.html