Infinium Mouse Methylation BeadChip GenomeStudio controls interpretation

To assess the performance of the Infinium Mouse Methylation BeadChip:

  • Create an Infinium Methylation project in the GenomeStudio 2011.1 Methylation Module using the Project Wizard as described in the GenomeStudio Methylation Module User Guide.

  • Infinium HD Methylation Controls are described in more detail in the Control Diagrams section (currently pages 96-103) of the Infinium HD Methylation Assay Reference Guide.

  • Infinium Methylation controls are viewed in GenomeStudio 2011.1 by navigating to Analysis > View Controls Dashboard.

  • The Mouse Methylation BeadChip includes control probes for both mouse and human targets.

Controls normalization for the Mouse Methylation BeadChip is not currently supported in GenomeStudio 2011.1 because the set of control probes used for normalization is specific to human sequences. BeadArray Controls Reporter (BACR) is not compatible with the Mouse Methylation Beadchip.

The following tables are a guide to evaluating the controls performance for the Mouse Methylation BeadChip. The expected intensity applies to only the targeted species, ie, probes labeled MUS apply only to mouse samples, and probes labeled HSA apply only to human samples.

Table 1: Sample-Independent Controls

Purpose

Name

Evaluate Green (GRN)

Evaluate Red (RED)

Expected Intensity

STAINING

STN-DNP-High

-

+

High

STAINING

STN-DNP-Bkg

-

+

Background

STAINING

STN-Biotin-High

+

-

High

STAINING

STN-Biotin-Bkg

+

-

Background

EXTENSION

EXT-G

+

-

High

EXTENSION

EXT-T

-

+

High

EXTENSION

EXT-A

-

+

High

EXTENSION

EXT-C

+

-

High

HYBRIDIZATION

HYB-Low

+

-

Low

HYBRIDIZATION

HYB-Medium

+

-

Medium

HYBRIDIZATION

HYB-High

+

-

High

TARGET REMOVAL

TRM-1

-

+

Low

TARGET REMOVAL

TRM-2

-

+

Low

Table 2: Sample-Dependent Controls

Purpose

Name

Species Targeted

Evaluate Green (GRN)

Evaluate Red (RED)

Expected Intensity

BISULFITE CONVERSION I

BS1-396C_MUS

Mus musculus

+

-

High

BISULFITE CONVERSION I

BS1-396U_MUS

Mus musculus

+

-

Background

BISULFITE CONVERSION I

BS1-140C_MUS

Mus musculus

+

-

Low

BISULFITE CONVERSION I

BS1-140U_MUS

Mus musculus

+

-

Background

BISULFITE CONVERSION I

BS1-409C_MUS

Mus musculus

-

+

High

BISULFITE CONVERSION I

BS1-409U_MUS

Mus musculus

-

+

Background

BISULFITE CONVERSION I

BS1-318C_MUS

Mus musculus

-

+

Medium

BISULFITE CONVERSION I

BS1-318U_MUS

Mus musculus

-

+

Background

BISULFITE CONVERSION I

BS1-317C_MUS

Mus musculus

-

+

Low

BISULFITE CONVERSION I

BS1-317U_MUS

Mus musculus

-

+

Background

BISULFITE CONVERSION I

BS1-C1_HSA

Homo sapiens

+

-

High

BISULFITE CONVERSION I

BS1-C2_HSA

Homo sapiens

+

-

Medium

BISULFITE CONVERSION I

BS1-C3_HSA

Homo sapiens

-

+

High

BISULFITE CONVERSION I

BS1-C4_HSA

Homo sapiens

-

+

High

BISULFITE CONVERSION I

BS1-C5_HSA

Homo sapiens

-

+

High

BISULFITE CONVERSION I

BS1-U1_HSA

Homo sapiens

+

-

Background

BISULFITE CONVERSION I

BS1-U2_HSA

Homo sapiens

+

-

Background

BISULFITE CONVERSION I

BS1-U3_HSA

Homo sapiens

-

+

Background

BISULFITE CONVERSION I

BS1-U4_HSA

Homo sapiens

-

+

Background

BISULFITE CONVERSION I

BS1-U5_HSA

Homo sapiens

-

+

Background

BISULFITE CONVERSION II

BS2-330_MUS

Mus musculus

-

+

High

BISULFITE CONVERSION II

BS2-505_MUS

Mus musculus

-

+

High

BISULFITE CONVERSION II

BS2-649_MUS

Mus musculus

-

+

High

BISULFITE CONVERSION II

BS2-1_HSA

Homo sapiens

-

+

High

BISULFITE CONVERSION II

BS2-3_HSA

Homo sapiens

-

+

High

BISULFITE CONVERSION II

BS2-4_HSA

Homo sapiens

-

+

High

SPECIFICITY I

SP1-PM1_HSA

Homo sapiens

+

-

High

SPECIFICITY I

SP1-PM2_HSA

Homo sapiens

+

-

High

SPECIFICITY I

SP1-PM3_HSA

Homo sapiens

+

-

High

SPECIFICITY I

SP1-MM1_HSA

Homo sapiens

+

-

Background

SPECIFICITY I

SP1-MM2_HSA

Homo sapiens

+

-

Background

SPECIFICITY I

SP1-MM3_HSA

Homo sapiens

+

-

Background

SPECIFICITY I

SP1-PM4_HSA

Homo sapiens

-

+

High

SPECIFICITY I

SP1-PM5_HSA

Homo sapiens

-

+

High

SPECIFICITY I

SP1-PM6_HSA

Homo sapiens

-

+

High

SPECIFICITY I

SP1-MM4_HSA

Homo sapiens

-

+

Background

SPECIFICITY I

SP1-MM5_HSA

Homo sapiens

-

+

Background

SPECIFICITY I

SP1-MM6_HSA

Homo sapiens

-

+

Background

SPECIFICITY II

SP2-1_HSA

Homo sapiens

-

+

High

SPECIFICITY II

SP2-2_HSA

Homo sapiens

-

+

High

SPECIFICITY II

SP2-3_HSA

Homo sapiens

-

+

High

NON-POLYMORPHIC

NPM-A_MUS

Mus musculus

-

+

High

NON-POLYMORPHIC

NPM-T_MUS

Mus musculus

-

+

High

NON-POLYMORPHIC

NPM-C_MUS

Mus musculus

+

-

High

NON-POLYMORPHIC

NPM-G_MUS

Mus musculus

+

-

High

NON-POLYMORPHIC

NPM-A_HSA

Homo sapiens

-

+

High

NON-POLYMORPHIC

NPM-T_HSA

Homo sapiens

-

+

High

NON-POLYMORPHIC

NPM-C_HSA

Homo sapiens

+

-

High

NON-POLYMORPHIC

NPM-G_HSA

Homo sapiens

+

-

High

NEGATIVE*

NEG-#

Homo sapiens, Mus musculus

+

+

Background

* Average intensity and standard deviation of 404 negative control probes.

Infinium Mouse Methylation BeadChip product files are available on the Infinium Mouse Methylation BeadChip Support Resources page.

For additional questions on Infinium controls evaluation, contact Illumina Technical Support at techsupport@illumina.com.

For any feedback or questions regarding this article (Illumina Knowledge Article #5666), contact Illumina Technical Support techsupport@illumina.com.

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