Luminex

The Biopolymers facility offers access to a Luminex 200 bead array instrument for running a host of XMAP based assays.

About Luminex Technology

• View slide based information about:


• Protein Technology


• Nucleic Acid Technology


• View a Flash presentation of the Technology Click here


• Luminex Assay General Information

The following information applies to many kits but each kit will come with its own protocols and specific instructions. The following information is provided as a general set of guidelines in thinking about your experiment set up. Please read and follow the manufacturers instructions for your specific kit.

Sample Layout
Each 96-well microplate can hold one 8-point standard curve and 40 samples run in duplicate. If you have two different media types or sample types that require a second standard curve on the same plate, then there is room for a total of 32 samples divided in the way that meets your needs.

Analyte Purchasing and Multiplexing
Please pay attention to each kit manufacturer's suggestions / restrictions on kit multiplexing. The kits must have compatible XMap bead panels and the tests themselves must have compatible reagents in terms of the conjugated molecules and kit reagents. Each manufacturer's kit will have this information. We do not recommend plexing different manufacturer's kits together unless you have confirmed they are compatible.

Sample Preparation
For cell culture supernatants, you will need to pre-clear your samples. There should be no cells present in the supernatant. You should also avoid multiple freeze-thaw cycles of your material. In general you will also need 15ml of your cell culture media for each sample type in order to dilute your samples and your standard curve dilutions as per the protocol you are using. Please note that we do not allow infectious or biohazardous material to be transported into the facility or to be assayed in our instrument.

Volumes
You will need a minimum of 50ul of your cell culture supernatant or serum sample. This volume will allow for duplicate wells. If the volume required cannot be provided, an increased dilution factor may be utilized but then the low-end sensitivity of your assay may be lost if there are only low levels of protein present.

• FlexmiR MicroRNA Panel

Using 10ug of human RNA, you can scan 319 microRNA in 4 hours.
The FlexmiR technology allows researchers to:

• Biotinylate the 3' ends of total RNA
• Hybridize the labeled RNA to Tm-normalized LNA capture probes that have been coupled to fluorescently dyed xMAP beads
• Tag the captured biotinylated miRNAs with streptavidin-phycoerythrin (SAPE)
• Analyze the samples on a Luminex 200.

You simply provide us with the RNA, and we return to you analyzed data of relative quantities of microRNAs are in your samples.

Rat/Mouse panels are also available.

For additional information about the technology, including a list of microRNA's in the panel, visit http://www.luminexcorp.com/microrna

• FlexmiR Select

Once you have selected targets based on a screen or through other methods, you may order custom pools of FlexmiR targets. These require far less RNA starting material thus reducing the cost of the assay.

For further information on FlexmiR Select, visit http://www.luminexcorp.com/microrna/flexmir_select.html

• FlexmiR Sample Submission Details

Samples need to be shipped to us frozen (on dry ice) in screw cap tubes.

For human pools, customer must provide at least 500ng/pool -2.5ug for the entire panel at 1ug/ul.

For human plus mouse/rat, customer must provide at least 500ng/pool-3.5ug for the entire panel at 1ug/ul.

FlexmiR Select requires 500ng based on one pool.

If you are able to submit 2 times the minimum amount, this will save time and cost in the event that a repeat is necessary.

Customer must provide a Bioanalyzer report of the samples, or provide an additional amount sufficient for Bioanalyzer (1-2ul of 50-500ng/ul). The RIN must be above 8.0 to be considered good. We will run with a lower RIN but we make no guarantee about data quality.

Back to top

Instrument use policy

The Luminex 200 is available on a shared instrumentation basis through the Biopolymers Facility. We require that you undergo training from one of our staff members on basic instrument use or provide proof of competency from prior instrument experience before using the instrument. Once you have been trained you may sign up for instrument use on our website.

What You Need:

You must provide your own pipetors, pipet tips, tubes, plates, foil and any other disposable or related equipment and materials. We strongly encourage users to conduct the majority of their Luminex bead experiment in their own lab and to perform only the instrument based data acquisition step at our facility. Many kits can take several hours to run and may require heated incubation steps or other steps for which we do not have the equipment available.

Important Note:

We currently do not allow any hazardous material (infectious virus or other biologically hazardous material) to be used in our Luminex Bead Array instrument.

Instrument Scheduling:

Instrument sign up is done through our website and web calendar system. It is important that you stick to your time as we must warm up the instrument prior to your arrival and perform a sanitizing wash cycle after you are done. If other users are scheduled to use the instrument and you are late it could disrupt their experiments and therefore you may be asked to reschedule.

Data Analysis:

The Bead View Software on the Luminex instrument computer is for data collection only and not for data analysis. We provide a separate computer in our shared computer facility with the bead view software that you can use to analyze your data. In addition, files must be transferred to your user folder as the beadview instrument computer will be purged of data weekly. Active protocols in the beadview instrument operating software will be maintained, protocols inactive for 90 days will be deleted.

What We Provide:

The Biopolymers Facility will calibrate the instrument and run the instrument start up protocols and procedures prior to your arrival. We will also run the instrument sanitization and shut down procedures after you are done. Our staff will also be available during your run to answer basic questions about operating the instrument should you need their assistance. Due to the very large number of assays available for this platform we do not have significant knowledge of them all so you should speak with the technical representative of the company from which you purchased your kit PRIOR to starting your experiment if you have any questions about the kit.

The following accessory equipment is available for Luminex experiments.

Back to top

Pricing

Instrument

FlexmiR MicroRNA Panel

FlexmiR Select

Back to top

©2004 Harvard Medical School