illumina Infinium HD Super Assay User manual

Infinium HD Super Assay
Reference Guide
Document # 11322427 v03
August 2019
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For Research Use Only. Not for use in diagnostic procedures.

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Document # 11322427 v03
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Infinium HD Super Assay Reference Guide

Revision History
Document Date Description of Change
Document # 11322427 v03 August 2019 Removed all references to WG# in barcode
information in regards to DNA plates and MSA
plates.
Added Barcode Numbers section to Tips and
Techniques to explain the new barcode
numbering scheme.
Document # 11322427 v02 June 2019 Removed instructions for 4x1 and 4x3
BeadChips.
Changed document name to Infinium HD Super
Assay Reference Guide .
Document updated to reflect latest style and
formatting standards.
Added Preparation and Storage of User-Supplied
Reagent section to consolidate reagent
preparation and use instructions.
Corrected storage temperatures for MA1, PB1,
RA1, XC3, and XC4.
Corrected reagents listed in first and last bullets
of About Reagents section in the Extend and
Stain introduction.
Reference to Experienced User Cards replaced
with Infinium HD Super Assay Checklist Manual
Protocol (document # 1000000080218) and
Infinium HD Super Assay Checklist Automated
Protocol (document # 1000000080220) which
provide current format of experienced user
instructions.
Added list of acronyms.
Consumables and equipment information moved
to Infinium Assay Lab Setup and Procedures
Guide (document # 11322460) .
Added Tip and Techniques section, includes:
• Tips, techniques, and general reagent storage
and preparation information previously
documented in protocol sections.
• Directions to seal plates before vortex and
centrifuge steps.
• BeadChip handling.
• Hybridization Chamber handling.
Document # 11322427 v01 November
2015
Corrected the hybridization oven temperature to
37°C when incubating the MSA1 plate and the
DNA.
Updated the total volume of PB1 required in
relation to the number of alignment fixtures in
use.
Document # 11322427 v03
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Infinium HD Super Assay Reference Guide

Document Date Description of Change
Part # 11294809 Rev. D May 2012 Added that up to 24 BeadChips can be washed
with 550 ml of PB1.
Part # 11294809 Rev. C June 2010 Update.
Part # 11294809 Rev. B June 2009 Update.
Part # 11294809 Rev. A November
2008
Minor revisions.
Part # 11294809 Rev. A May 2008 Initial release.
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Infinium HD Super Assay Reference Guide

Table of Contents
Chapter 1 Overview 1
Introduction to Infinium HD Super Assay 1
Important Note 2
Additional Resources 2
DMAPFiles 2
Preparation and Storage of User-Supplied Reagents 2
Tips and Techniques 3
Acronyms 5
Illumina LIMS 5
No Illumina LIMS 6
Tracking Tools 6
Chapter 2 Manual Protocol 7
Introduction 7
Infinium HD Super Manual Workflow 7
Amplify DNA 9
Incubate DNA 10
Fragment DNA 10
Precipitate DNA 11
Resuspend DNA 13
Hybridize to BeadChip 14
Wash BeadChips 19
Extend and Stain BeadChips 25
Image BeadChip 34
Illumina GenomeStudio 35
Chapter 3 Automated Protocol 37
Introduction 37
Infinium HD Super Automated Workflow 37
Amplify DNA 39
Incubate DNA 41
Fragment DNA 41
Precipitate DNA 42
Resuspend DNA 44
Hybridize to BeadChip 46
Wash BeadChips 53
Extend and Stain BeadChips 58
Image BeadChip 68
Illumina GenomeStudio 69
Technical Assistance 70

Chapter 1 Overview
Overview
Introduction to Infinium HD Super Assay 1
Important Note 2
Additional Resources 2
DMAPFiles 2
Preparation and Storage of User-Supplied Reagents 2
Tips and Techniques 3
Acronyms 5
Illumina LIMS 5
No Illumina LIMS 6
Tracking Tools 6
Introduction to Infinium HD Super Assay
The Illumina Infinium HD Super Assay is an evolution for DNA analysis, by streamlining sample preparation and
enabling high multiplexing. Using Infinium I and Infinium II probe designs and a dual-color channel approach,
the Infinium HD Super Assay enables the DNA analysis of up to several million of SNPs and CNV markers per
sample.
The Infinium HD Super Assay accomplishes this unlimited multiplexing by combining whole-genome
amplification (WGA) sample preparation with direct, array-based capture, and enzymatic scoring of the SNP
loci. Locus discrimination or copy number variation (CNV) determination comes from a combination of high
bead type representation per feature, sequence-specific hybridization capture and array-based, single-base
primer extension. In the case of the Infinium II probe design, the 3' end of the primer is positioned directly
adjacent to the SNP site, or if a nonpolymorphic probe, directly adjacent to the nonpolymorphic site. With the
Infinium I probe design, the 3' end of the primer overlaps with the SNP site. If there is a perfect match,
extension occurs and signal is generated. If there is a mismatch, extension does not occur and no signal is
generated.
Allele-specific single base extension of the primer incorporates a biotin nucleotide or a dinitrophenyl labeled
nucleotide. C and G nucleotides are biotin labeled; A and T nucleotides are dinitrophenyl labeled. Signal
amplification of the incorporated label further improves the overall signal-to-noise ratio of the assay.
The Infinium HD Super Assay offers:
uHigh multiplexing
uHigh call rate and accuracy
uUnlimited genomewide marker selection
uSingle tube amplification, single chip, no PCR
uMinimal risk of carryover contamination
uLow DNA input—200 ng per sample
uWalk-away automation using Tecan Genesis or Freedom EVO Robots and Tecan GenePaint system
uCompatibility with Illumina iScan and HiScanSystem
uMultiple-sample BeadChip format
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Important Note
Before using the procedures in this guide, read the Infinium Lab Setup and Procedures Guide . The Setup
and Procedures guide explains how to equip and run an Infinium HD Super Assay laboratory, including
information on the following topics:
uPrevention of amplification product contamination
uSafety precautions
uConsumables and equipment to purchase in advance
uStandard lab procedures
uRobot use for automation
uPreparation for BeadChip imaging
uSystem Controls
uSystem maintenance
uTroubleshooting
The instructions apply equally to all Infinium BeadChips provided by Illumina™. All Infinium HD Super
documentation assumes that you have already set up the laboratory space and that you are familiar with the
standard procedures and safety precautions.
Additional Resources
Visit the Infinium HD Super Assay support page on the Illumina website for documentation, software
downloads, training resources, and information about compatible Illumina products.
The following documentation is available for download from the Illumina website.
Resource Description
Infinium HD Super Assay Checklist Automated Protocol
(document # 1000000080220)
Provides a checklist for experienced users of the Infinium
HD Super Assay when performing the automated protocol.
Infinium HD Super Assay Checklist Manual Protocol
(document # 1000000080218)
Provides a checklist for experienced users of the Infinium
HD Super Assay when performing the manual protocol.
Infinium Lab Setup and Procedures Guide (document
#11322460)
Describes how to set up an Infinium HD Super lab
including reagents, consumables, and equipment to
purchase in advance, and best practices for lab operation.
Infinium Consumables and Equipment List (document #
1000000084294)
Provides an interactive checklist of Illumina-provided and
user-provided consumables and equipment.
DMAPFiles
Before run setup, download the DMAP files for the arrays, and prepare a sample sheet. Use the Decode File
Client to download the DMAP files.
Preparation and Storage of User-Supplied Reagents
uMaintain a first in, first out (FIFO) system for reagents. Rotate the stock of the remaining reagents to avoid
using expired reagents.
uInfinium HD Super kits contain reagents in exact quantities needed for the assay. Measure reagents
carefully to avoid shortages.
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uUse fresh reagents for each batch of plates, and empty reservoirs between batches.
uTo minimize errors when preparing user-supplied reagents, prepare large batches of 0.1 N NaOH and
95% Formamide/1 mM EDTA using the following guidance.
Preparing Batches of 0.1 N NaOH
Prepare fresh 0.1 N NaOH in large batches. Divide batches into 15 ml or 50ml sealed tubes.
Store the sealed tubes for 6 months at 2°C to 8°C and use the stored 0.1 NNaOHas needed. Use the 0.1
NNaOHthe same day you open the tube, and discard any unused amounts.
Preparing Batches of 95% Formamide/1 mM EDTA
Prepare the 95%formamide/1mM EDTA mixture in large batches. Divide batches into 15ml or 50 ml sealed
tubes.
Store the sealed tubes for up to 6 months at -25°C to -15°C and use the stored mixture as needed. Use the
mixture the same day you open the tube, and discard any unused amounts.
Use Fresh RA1 Reagent for Each Step
Use fresh RA1 for each step in the assay where it is required. RA1 that has been stored properly and has not
been dispensed for use in either the XStain or Resuspension step is considered fresh RA1. After RA1 has
been exposed to room temperature air for extended periods of time, it is no longer fresh.
To make best use of RA1, only pour the amount needed for the current step. If you plan to perform additional
assay steps requiring RA1 the same day, leave the remaining thawed reagent in the original, closed bottle at
room temperature until it is needed. Follow the standard RA1 storage procedures described in this guide for
next-day processing and prolonged storage conditions.
Store PB1
For Infinium HD Super workflows do the following.
uStore PB1 at room temperature.
uKeep PB1 for up to 3 months.
Tips and Techniques
Unless a safe stopping point is specified, proceed immediately to the next step.
Avoid Cross-Contamination
When adding or transferring samples, change tips between each sample .
Measure Volumes Carefully
When measuring volumes of reagent to put in reservoirs, make sure that the measurements are exactly as
specified in the instructions. Do not estimate volumes.
HD Glass Back Plates
uFor optimal performance, use HD glass back plates that are free of chips and cracks along the beveled
edge and the long edges. Chipped or cracked HD glass back plates risk volume leakage and affect
overall performance.
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uClean HD glass back plates according to the procedures in the Infinium Assay Lab Setup and Procedures
Guide (Document # 11322460) .
uA 10% bleach soak is required weekly, or every seven uses, to maintain glass performance.
Barcode Numbers
uBarcode serial numbers used for workflow enforcement and positive sample tracking on MSA plates,
DNA plates, and reagents use a numbering pattern of two alphanumeric characters followed by seven
random numbers.
Sealing the Plate
uAlways seal plates before the vortex and centrifuge steps in the protocol.
uOrient sealing mats so that the A1 on a cap matches the A1 on the plate.
uMake sure that all 96 caps are securely seated in the wells to prevent evaporation and spills, which
introduce variability and cross-contamination.
uRemove sealing mats slowly and carefully to prevent splashing, and then set aside upside-down in a
safe location.
uWhen returning a sealing mat to a plate, make sure that the orientation is correct.
Heat Sealer
uIf you are using the ALPS 50 V model heat sealer, set it to 165°C and 2.5 seconds.
Pipetting
uMake sure that pipettes are properly calibrated, cleaned, and decontaminated.
uDispense slowly and carefully to prevent turbulence in the plate wells and flow-through chambers.
uUse a multichannel pipette whenever possible.
Centrifugation
uWhen centrifuging plates or BeadChips, place a balance plate or rack with BeadChips opposite each
plate or rack being centrifuged. Make sure that the weights are as similar as possible.
Washing and Coating BeadChips
Perform the following steps before starting the wash and coat process:
uPlace wash dish covers on wash dishes when not in use.
uClean wash dishes with low-pressure air to remove particulates before use.
uWash tube racks and wash dishes thoroughly before and after use. Rinse with DI H2O. Place them
upside down on wash rack to dry.
uPrepare an additional clean tube rack that fits the internal dimensions of the vacuum desiccator. Allow
one rack per eight BeadChips.
Handling Hybridization Chambers
uKeep the chamber lids and bases together. Adopt a labeling convention that pairs each chamber base
with the original lid.
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uRegularly check lid-base pairs to make sure that they fit securely. Also check hinges for signs of abnormal
wear or loose fittings. An airtight seal requires that the hinges have adequate clamping strength.
uRecord which hybridization chamber was used for each BeadChip. If sample evaporation or other
processing anomalies occur, investigate the appropriate hybridization chambers.
uWhen the hybridization chamber inserts contain BeadChips, keep them steady and level when lifting or
moving.
uAvoid shaking, and always keep parallel to the lab bench.
uDo not hold by the sides near the sample inlets.
Rocker Platform [Optional]
If you are using an Illumina Hybridization Oven with a rocker platform, follow these guidelines:
uIf 7 to12 hybridization chambers (Hyb Chambers) are placed in 1 hybridization oven, the rocker must be
removed before beginning the Hybridize to BeadChip section.
uIf 1 to 6 Hyb Chambers are placed in 1 hybridization oven, removing the rocker platform before beginning
the Hybridize to BeadChip section is optional.
uRemoving the rocker platform is optional for plate incubation in the Amplify DNA, Incubate DNA,
Fragment DNA, Precipitate DNA, and Resuspend DNA sections.
Acronyms
Acronym Definition
EDTA Ethylenediaminetetraacetic acid
EtOH Ethanol
ATM Anti-Stain Two-Color Master Mix
FMS Fragmentation solution
MA1 Multi-Sample Amplification 1 Mix
MA2 Multi-Sample Amplification 2 Mix
MSM Multi-Sample Amplification Master Mix
PB1 Reagent used to prepare BeadChips for hybridization
PB2 Humidifying buffer used during hybridization
PM1 Precipitation solution
RA1 Resuspension, hybridization, and wash solution
STM Superior Two-Color Master Mix
TEM Two-Color Extension Master Mix
XC1 XStain BeadChip solution 1
XC2 XStain BeadChip solution 2
XC3 XStain BeadChip solution 3
XC4 XStain BeadChip solution 4
Illumina LIMS
If you are using the automated protocol with Illumina LIMS, follow these guidelines:
uAt each step in the protocol, make sure that the Use Barcodes checkbox is selected.
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uThe barcode must be on the right side of the plate. Make sure the barcode label fits between the
notches and does not cover the holes on the top of the plate.
Figure 1 Correctly Placed Barcodes
uWhen prompted, enter the number of samples, plates, or BeadChips, and then select OK.
uEach time you select Run to start a new process, you are prompted to log in to LIMS.
uIf prompted to select the project and the batch IDor DNAplate, do one of the following:
uSelect your current project. The available batches appear in the Sample Batch ID pane. Select a
batch to see the associated DNA plate appear in the DNA Plates pane.
uUse Search to search for a specific Batch ID or DNA plate.
uSome protocol steps require verification in the LIMS HTML user interface before you can begin
processing on the robot.
uIf verification is successful, a blue confirmation message appears at the top of the window.
uIf the verification fails, a red error message appears at the top of the window. Do not proceed.
Instead, follow these steps to troubleshoot the problem:
a Select the Reports tab in the upper-right corner.
b In the left pane, select Tracking Reports | Get Queue Status.
c Scan the plate barcode, and select Go.
d Note which step the plate is queued to run, and proceed with that step.
e If the error message indicates the BeadChip did not accession into the system, accession it and
repeat the verification step.
f If the BeadChip is not the right type for the batch, accession the correct type and repeat the
verification step.
No Illumina LIMS
If you are using the automated protocol without Illumina LIMS do the following:
uAt each step in the protocol, make sure that the Use Barcodes checkbox is cleared.
uAt each step in the protocol, you are prompted to enter the number of samples, plates, or BeadChips.
Enter the requested information, and then select OK.
Tracking Tools
Illumina provides the following tools for sample tracking and guidance in the lab:
uChecklists to guide you through the protocols. There is a checklist for each protocol, manual and
automated.
uLab Tracking Form to map DNA samples to BeadChips and record the barcode of each reagent and
plate used in the protocol. Record operator information, and start and stop times.
uSample sheet template to record information about your samples for later use in data analysis.
These documents are available for printing and reference at www.illumina.com/documentation.
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Chapter 2 Manual Protocol
Introduction 7
Infinium HD Super Manual Workflow 7
Amplify DNA 9
Incubate DNA 10
Fragment DNA 10
Precipitate DNA 11
Resuspend DNA 13
Hybridize to BeadChip 14
Wash BeadChips 19
Extend and Stain BeadChips 25
Image BeadChip 34
Illumina GenomeStudio 35
Introduction
This section describes pre- and post-amplification manual laboratory protocols for the Infinium HD Super
Assay. Follow the protocols in the order shown.
Infinium HD Super Manual Workflow
The following figure graphically represents the Infinium HD Super Assay manual workflow for 24 BeadChips.
These protocols describe the procedure for preparing 96 DNA samples.
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Figure 2 Infinium HD Super Manual Workflow
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Amplify DNA
This step adds the DNA samples to the plates. The samples are denatured and neutralized to prepare them
for amplification.
Consumables
uMA1 (2 tubes)
uMA2 (2 tubes)
uMSM (2 tubes)
u0.1N NaOH (15 ml)
u96-well 0.8 ml microplate (midi) (1 plate)
uDNA plate with 48 or 96 DNA samples (50 ng/μl) (midi or TCY) (1 plate)
uCap mats
Preparation
1 Preheat the Illumina Hybridization Oven in the post-amp area to 37°C and allow the temperature to
equilibrate.
2 Prepare the following consumables:
Item Storage Instructions
DNA -25°C to -15°C Thaw at room temperature. DNA must be 50 ng/µl, resuspended in TE (10 mM Tris,
1mM EDTA).
MA1 Room
temprature
Thaw at room temperature. Invert 10 times to mix, and then pulse centrifuge to
eliminate bubbles and collect reagent at the bottom of the tube.
MA2 -25°C to -15°C Thaw at room temperature. Invert 10 times to mix, and then pulse centrifuge to
eliminate bubbles and collect reagent at the bottom of the tube.
MSM -25°C to -15°C Thaw at room temperature. Invert 10 times to mix, and then pulse centrifuge to
eliminate bubbles and collect reagent at the bottom of the tube.
3 Apply an MSA1 barcode label to a new 0.8 ml microplate (midi).
Procedure
1 If you do not already have a DNA plate, add DNA into either of the following:
uMidi plate: 20 µl to each DNA well
uTCY plate: 10 µl to each DNA well
Apply a barcode label to the new DNA plate.
2 Add 20µl MA1 into the MSA1 plate wells.
3 Transfer 4µl DNA sample from the DNA plate to the corresponding wells in the MSA1 plate.
4 Add 4µl 0.1N NaOH into each well of the MSA1 plate.
Refer to the following figure throughout the Make MSA1 process.
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5 Seal the MSA1 plate with the 96-well cap mat.
uOrient the mat so that A1 on the cap matches A1 on the plate.
uMake sure that all 96 caps are securely seated in the wells.
6 Vortex the plate at 1600rpm for 1minute, and then pulse centrifuge at 280× g.
7 Incubate at room temperature for 10minutes.
8 Remove the cap mat and set aside upside down in a safe location.
9 Add 68µl MA2 into each well of the MSA1 plate.
10 Add 75µl MSM into each well of the MSA1 plate.
11 Reseal with the cap mat using the original orientation.
12 Vortex at 1600 rpm for 1 minute, and then pulse centrifuge at 280 × g.
NOTE
Perform the remaining protocol steps in the post-amplification area.
Incubate DNA
This step uniformly amplifies the genomic DNA, generating a sufficient quantity of each individual DNA sample
to be used in the Infinium HD Super Assay.
1 Incubate the MSA1 plate in the Illumina Hybridization Oven for 20–24 hours at 37°C.
Fragment DNA
This step enzymatically fragments the DNA. An endpoint fragmentation is used to prevent overfragmentation.
Consumables
uFMS (2 tubes)
uCap mat
Preparation
1 Preheat the heat block with the midi plate insert to 37°C.
2 Prepare the following consumable.
Item Storage Instructions
FMS -25°C to -15°C Thaw to room temperature. Invert 10 times to mix.
3 Remove the MSA1 plate from the Illumina Hybridization Oven.
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Procedure
1 Pulse centrifuge the MSA1 plate at 280 × g.
2 Carefully remove the cap mat.
3 Add 50µl FMS to each well of the MSA1 plate.
4 Reseal with the cap mat using the original orientation.
5 Vortex at 1600 rpm for 1 minute, and then centrifuge the plate at 280 × g.
6 Incubate on the preheated heat block for 1 hour.
If you are continuing, you can leave the plates on the heat block until you have completed preparation for
the next step, no longer than 2hours.
SAFE STOPPING POINT
If you are stopping, seal the plate, and store at -25°C to -15°C for up to 24 hours.
Precipitate DNA
This step uses 100 % 2-propanol and PM1 to precipitate the DNA.
Consumables
u100% 2-propanol
uPM1(2 tubes)
uCap mat
Preparation
1 Do one of the following:
uIf proceeding immediately from Fragment DNA , leave the MSA1 plate on the heat block until
preparation is complete.
uIf the MSA1 plate was stored at -25°C to -15°C, thaw at room temperature, pulse centrifuge
at280×g, and preheat the heat block to 37°C.
2 Prepare the following consumable.
Item Storage Instructions
PM1 2°C to 8°C Thaw to room temperature and invert 10 times to mix.
3 Remove the cap mat.
Procedure
1 Add 100 μl PM1 to each well of the MSA1 plate.
2 Reseal with the cap mat using the original orientation.
3 Vortex the plate at 1600rpm for 1minute.
4 Incubate on the preheated heat block for 5minutes.
5 Pulse centrifuge at 280 × g for 1 minute.
6 Set the centrifuge at 4°C in preparation for the next centrifuge step.
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7 Remove and discard the cap mat.
8 Add 300 μl 100% 2-propanol to each sample well.
9 Carefully seal with a new, dry cap mat. Avoid shaking the plate until the cap mat is seated.
10 Invert the plate 10 times to mix.
11 Incubate in a refrigerator set at 4°C for 30minutes.
12 Centrifuge at 3000× g at 4°C for 20minutes.
13 Immediately remove the plate from the centrifuge.
uWhen centrifuging is complete, proceed immediately to avoid dislodging the blue pellets.
uIf a delay occurs, repeat the 20minute centrifuge.
14 Make sure that a blue pellet is present in the bottom of each sample well.
15 Remove and discard the cap mat.
16 Hold the plate over an absorbent pad and do as follows.
a Quickly invert to decant the supernatant.
b Drain liquid onto the absorbent pad, and then smack the plate down on a dry area of the pad.
17 Keeping the plate inverted, firmly tap until all wells are free of liquid (~1 minute). Do not allow supernatant
to pour in to other wells.
18 Place the uncovered, inverted plate on the tube rack for 1 hour at room temperature to air-dry the
pellets.
19 Make sure that a blue pellet is still present in the bottom of each sample well.
20 Keeping the plate inverted, use a Kimwipe to remove any residual alcohol draining from the wells of the
plate or remaining on the surface of the plate.
CAUTION
Do not overdry the pellets. Pellets that are overdried are difficult to resuspend and can lead to poor data
quality.
SAFE STOPPING POINT
If you are stopping, seal the plate, and store at -25°C to -15°C for up to 24 hours.
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Resuspend DNA
This step uses RA1 to resuspend the precipitated DNA.
Consumables
uRA1 (46 µl per well)
uFoil heat seals
About Reagents
uDispense only the volume necessary for each step. The Extend and Stain BeadChips procedure also
requires RA1.
uUse fresh RA1 for each step that requires it. Properly stored RA1 that has not been dispensed for this
resuspension step or the extend and stain step is considered fresh.
uRA1 might form visible precipitate or crystals. Before each use, hold in front of a light and inspect. Invert
several times to redissolve the solution as needed.
WARNING
This protocol uses an aliphatic amide that is a probable reproductive toxin. Personal injury can occur through
inhalation, ingestion, skin contact, and eye contact. For more information, consult the material data safety
sheet for this assay at www.illumina.com/sds. Dispose of containers and any unused contents in
accordance with the governmental safety standards for your region.
Preparation
1 If the MSA1 plate was stored at -25°C to -15°C, thaw to room temperature, then remove cap mat.
2 Prepare the following consumable.
Item Storage Instructions
RA1 -25°Cto-15°C Warm to room temperature in a 20°C to 25°C water bath. Mix to dissolve any remaining
crystals.
3 Preheat the Illumina Hybridization Oven to 48°C.
4 Preheat the heat sealer for at least 20 minutes before use.
Procedure
1 Add 46µl RA1 to each well of the MSA1 plate.
2 With the dull side facing down, apply a foil heat seal to the plate. Hold the heat sealer sealing block down
firmly and evenly for 5seconds.
3 Incubate in the Illumina Hybridization Oven for 1 hour.
4 Vortex at 1800rpm for 1minute.
5 Pulse centrifuge at 280 × g.
6 Repeat steps 4and 5as needed to resuspend the pellets.
7[Optional] Unless you are stopping, you can set aside the MSA1 plate for up to 1 hour before proceeding.
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SAFE STOPPING POINT
If you are stopping, seal the plate, and store at -25°C to -15°C for up to 24 hours.
Hybridize to BeadChip
This step dispenses the fragmented, resuspended DNA onto BeadChips. Incubation then hybridizes each
DNA sample to a section of the BeadChip.
Consumables
u100 % EtOH (330 ml)
uPB2 (2 tubes)
uXC4
About Reagents
uKeep XC4 in the original bottle until you are ready to use it.
uEach XC4 bottle contains sufficient reagent to process up to 24 BeadChips.
uUse resuspended XC4 at room temperature.
Preparation
1 If frozen, thaw the MSA1 plate at room temperature, and then pulse centrifuge at 280 × g.
2 Preheat the heat block to 95°C.
3 Preheat the Illumina Hybridization Oven to 48°C.
Procedure
Denature DNA
1 Place the MSA1 plate on the preheated heat block for 20 minutes to denature the DNA.
2 Cool the MSA1 plate on the benchtop at room temperature for 30minutes.
3 Pulse centrifuge at 280 × g.
Assemble the Hybridization Chambers
Assemble one chamber for every four BeadChips by following the steps in this section.
1 Place the hybridization chambers, hybridization chamber gaskets, and hybridization chamber inserts on
the benchtop.
Document # 11322427 v03
For Research Use Only. Not for use in diagnostic procedures.
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Infinium HD Super Assay Reference Guide