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Biopolymers Facility at Harvard Medical School
The Biopolymers facility offers access
to a Luminex 200 bead array instrument for running a host of XMAP based assays.
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.
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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.
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.
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 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.
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.
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.
Branson 1.9L 1510 ultrasonic cleaner.
(View Picture)
This deluxe model features digital heat and timer controls. Specifications Overall Size 10" x 12" x 11.5" Tank Size 6" x 5.5" x 4" Weight 7 lbs.
Barnstead Orbital Plate Rotator
(View Picture)
6.5" X 6.5" Platform, 0 to 240 rpm platform speed with 60 minute timer.
QIAvac 96 Vacuum Manifold
(View Picture)
QIAvac 96 is designed for efficient processing of 96-well filter plates.
The 96-well purification module sits in the QIAvac 96 top plate, allowing
samples and buffers to be efficiently drawn through the wells. The versatile
QIAvac base holds an additional module, a waste tray, or collection vessels
such as microtubes or a 96-well microplate during processing. This allows
samples to be drawn from one module to the next, avoiding time-consuming labeling
and transfer steps.
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| Item |
Harvard |
Non-Harvard/Academic |
Commercial |
| Initial training per lab member |
$100.00 |
$109.58 |
$120.00 |
| Includes instrument use training. |
| Instrument runs per 96-well plate |
$50.00 |
$54.79 |
$60.00 |
| Full charge whether plate is full or not |
| Staff run experiment per 96-well plate |
$75.00 |
$82.18 |
$90.00 |
| Full charge whether plate is full or not |
| Data Distribution |
Included. |
Included. |
Included. |
| Data will be transferred to your ftp user folder |
| Data Analysis Software |
Included. |
Included. |
Included. |
| Use of Beadview software in our shared computer facility is currently included for users conducting Luminex experiments in our facility. |
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