For Research Use Only. Not for use in diagnostic procedures.

DISCOVERYDETECTION

SeqCap EZ Cardiology PanelsAnalyze genetic mutations associated with the research of hereditary cardiac disorders

SeqCap EZ Cardiology Panels enable researchers to detect variants in genes that are most commonly associated with hereditary cardiac disorders. Developed using guidance from publications, literature, and Roche Sequencing Solutions’ scientific experts, SeqCap EZ Cardiology Panels are optimized for use with HyperCap Workflow, which integrates KAPA library preparation and SeqCap EZ target enrichment products in a single streamlined workflow.

Panels Description• SeqCap EZ Cardiomyopathy Panel: 76 gene (372 kb) panel for research on genes commonly associated with

heart muscle disorders

• SeqCap EZ Channelopathy and Arrhythmias Panel: 54 gene (203 kb) panel for research on genes involved in ion channel defects and irregular heartbeat

• SeqCap EZ Sudden Cardiac Death Panel: 140 gene (610 kb) panel for research on genes associated with Cardiomyopathies, Arrhythmias, Channelopathies, Marfan’s Syndrome, and Loeys-Dietz Syndrome

Features and Benefits• Panel breadth: Focused and comprehensive panels to encompass different research needs

• Improved uniformity: Enabling more even target coverage and increased sequencing efficiency

• Expert-driven content: Comprehensive gene coverage for cardiology research enabling rare variant detection

• Single vendor and support: Compatible with the Hypercap Workflow, convenient one-stop shop for ordering, service, and support for a complete sample prep solution

2Data on file.For Research Use Only. Not for use in diagnostic procedures.

Performance MetricsSeqCap EZ Cardiomyopathy Panel

• Comprehensive coverage of 76 genes from CCDS and ClinVar databases for cardiomyopathy research

• Achieve uniform sequencing depth with a low Fold 80 base penalty

• Complete coverage at 20X and nearly complete coverage at 50X when this research panel is sequenced to an average depth of 250X

(A) Efficiency of sequencing. Percent of mapped non-duplicate reads on-target is the percent of mapped, de-duplicated reads overlapping the target region by at least 1 bp, with no padding or buffer. The percent of bases in padded targets is the percent of captured and sequenced bases which fall within 250 bp of the primary target regions of the panel. (B) Uniformity of sequencing. Fold 80 base penalty is a measure of uniformity produced by the Picard tools, and is the fold additional sequencing required to bring 80% of bases to the mean coverage. This is a measure of sequence depth uniformity—lower is better with the best theoretical value equal to 1. Zero coverage regions are excluded. (C) Coverage for variant calling. Bars indicate complete coverage at 20X for all panels, and nearly complete coverage at 50X, when each panel is sequenced to an average depth of 250X. (D) Depth of coverage versus number of sequencing reads. Replicate captures were performed for each cardiology research panel. Eight different HapMap samples were prepared using KAPA HyperPrep Library Preparation Kit following the SeqCap EZ HyperCap Workflow User’s Guide v2.0. Captures were 8-plexed (using 125 ng of each sample library). Each capture was sequenced using an Illumina® MiSeq® sequencing instrument (MiSeq Reagent Kit v2; 2 x 101 bp) Reads were sampled at intervals of 250,000 reads, and the mean, de-duplicated depth of coverage was determined at each sampling level. (A, B, and C) Replicate captures were performed for each cardiology research panel. Eight different HapMap samples were prepared using KAPA HyperPrep Library Preparation Kit following the SeqCap EZ HyperCap Workflow User’s Guide v2.0. Captures were 8-plexed (using 125 ng of each sample library). Each capture was sequenced using an Illumina MiSeq sequencing instrument (MiSeq Reagent Kit v2; 2 x 101 bp) Data was subsampled to an average coverage depth of 250X.

SeqCap EZ Cardiomyopathy Panel

Panel Size 76 genes; 372 kb

Disorders Covered by Research Panel

Hypertrophic Cardiomyopathy, Arrhythmogenic Right Ventricular Cardiomyopathy, Dilated Cardiomyopathy, Left Ventricular Non-Compaction Cardiomyopathy, and Restrictive Cardiomyopathy

Product Type SeqCap EZ Share Prime Choice

Internal Reference # 4000034930

Design Name 171129_HG38_CardioP1_REZ_HX3

Replicate 1 Replicate 2

Percent of reads on-target and percent of bases in padded target panel

Average of % mapped non-duplicate reads on-target (primary) Average of % bases in padded target (capture)

61.8

73.4 74.0

87.3

0

10

20

30

40

50

60

70

80

90

100

Replicate 1 Replicate 2

Fold 80 base penalty

Average of Fold 80 base penalty

3.0

2.5

1.5

2.0

1.0

0.5

0

1.361.38

Mean coverage vs reads

Replicate 1 Replicate 2

0

50

100

150

200

250

300

350

400

0 500000 1000000 1500000 2000000 2500000 3000000 3500000Replicate 1 Replicate 2

Percent of bases covered at 20X and 50X

Average of % bases ≥20 Average of % bases ≥50

100 99.90 100 99.96

0

20

40

60

80

100

120

A B

C D

3Data on file.For Research Use Only. Not for use in diagnostic procedures.

SeqCap EZ Channelopathy and Arrhythmias Panel

• Comprehensive coverage of 54 genes from CCDS and ClinVar databases for channelopathy and arrhythmias research

• Achieve uniform sequencing depth with a low Fold 80 base penalty

• Complete coverage at 20X and nearly complete coverage at 50X when this research panel is sequenced to an average depth of 250X

SeqCap EZ Channelopathy and Arrhythmias Panel

Panel Size 54 genes; 203 kb

Disorders Covered by Research Panel

Long QT 1-7, Cathecholaminergic Polymorphic Ventricular Tachycardia, Idiopathic VT (regional), Brugada Syndrome, Arrythmogenic Right Ventricular Cardiomyophathy, Sick Sinus Syndrome/Atrial Standstill, Short QT Syndrome, and Familial Atrial Fibrillation

Product Type SeqCap EZ Share Prime Choice

Internal Reference # 4000034940

Design Name 171129_HG38_CardioP2_REZ_HX3

(A) Efficiency of sequencing. Percent of mapped non-duplicate reads on-target is the percent of mapped, de-duplicated reads overlapping the target region by at least 1 bp, with no padding or buffer. The percent of bases in padded targets is the percent of captured and sequenced bases which fall within 250 bp of the primary target regions of the panel. (B) Uniformity of sequencing. Fold 80 base penalty is a measure of uniformity produced by the Picard tools, and is the fold additional sequencing required to bring 80% of bases to the mean coverage. This is a measure of sequence depth uniformity—lower is better with the best theoretical value equal to 1. Zero coverage regions are excluded. (C) Coverage for variant calling. Bars indicate complete coverage at 20X for all panels, and nearly complete coverage at 50X, when each panel is sequenced to an average depth of 250X. (D) Depth of coverage versus number of sequencing reads. Replicate captures were performed for each cardiology research panel. Eight different HapMap samples were prepared using KAPA HyperPrep Library Preparation Kit following the SeqCap EZ HyperCap Workflow User’s Guide v2.0. Captures were 8-plexed (using 125 ng of each sample library). Each capture was sequenced using an Illumina® MiSeq® sequencing instrument (MiSeq Reagent Kit v2; 2 x 101 bp). Reads were sampled at intervals of 250,000 reads, and the mean, de-duplicated depth of coverage was determined at each sampling level. (A, B, and C) Replicate captures were performed for each cardiology research panel. Eight different HapMap samples were prepared using KAPA HyperPrep Library Preparation Kit following the SeqCap EZ HyperCap Workflow User’s Guide v2.0. Captures were 8-plexed (using 125 ng of each sample library). Each capture was sequenced using an Illumina MiSeq sequencing instrument (MiSeq Reagent Kit v2; 2 x 101 bp). Data was subsampled to an average coverage depth of 250X.

Replicate 1 Replicate 2

Percent of reads on-target and percent of bases in padded target panel

Average of % mapped non-duplicate reads on-target (primary) Average of % bases in padded target (capture)

66.4

84.7

69.8

88.8

0

10

20

30

40

50

60

70

80

90

100

Replicate 1 Replicate 2

Fold 80 base penalty

Average of Fold 80 base penalty

3.0

2.5

1.5

2.0

1.0

0.5

0

1.38 1.37

Mean coverage vs reads

Replicate 1 Replicate 2

0

50

100

150

200

250

300

400

350

450

0 500000 1000000 1500000 2000000 2500000Replicate 1 Replicate 2

Percent of bases covered at 20X and 50X

Average of % bases ≥20 Average of % bases ≥50

100 100 100 100

0

20

40

60

80

100

120

A B

C D

4Data on file.For Research Use Only. Not for use in diagnostic procedures.

SeqCap EZ Sudden Cardiac Death Panel

• Comprehensive coverage of 140 genes from CCDS and ClinVar databases for sudden cardiac death research

• Achieve uniform sequencing depth with a low Fold 80 base penalty

• Complete coverage at 20X and nearly complete coverage at 50X when this research panel is sequenced to an average depth of 250X

SeqCap EZ Sudden Cardiac Death Panel

Panel Size 140 genes; 610 kb

Disorders Covered by Research Panel Panel 1 disorders + Panel 2 disorders plus other thoracic aortic aneurysms and dissections, Marfan’s Syndrome, and Loeys-Dietz Syndrome

Product Type SeqCap EZ Share Prime Choice

Internal Reference # 4000034950

Design Name 171129_HG38_CardioP3_REZ_HX3

(A) Efficiency of sequencing. Percent of mapped non-duplicate reads on-target is the percent of mapped, de-duplicated reads overlapping the target region by at least 1 bp, with no padding or buffer. The percent of bases in padded targets is the percent of captured and sequenced bases which fall within 250 bp of the primary target regions of the panel. (B) Uniformity of sequencing. Fold 80 base penalty is a measure of uniformity produced by the Picard tools, and is the fold additional sequencing required to bring 80% of bases to the mean coverage. This is a measure of sequence depth uniformity—lower is better with the best theoretical value equal to 1. Zero coverage regions are excluded. (C) Coverage for variant calling. Bars indicate complete coverage at 20X for all panels, and nearly complete coverage at 50X, when each panel is sequenced to an average depth of 250X. (D) Depth of coverage versus number of sequencing reads. Replicate captures were performed for each cardiology research panel. Eight different HapMap samples were prepared using KAPA HyperPrep Library Preparation Kit following the SeqCap EZ HyperCap Workflow User’s Guide v2.0. Captures were 8-plexed (using 125 ng of each sample library). Each capture was sequenced using an Illumina® MiSeq® sequencing instrument (MiSeq Reagent Kit v2; 2 x 101 bp). Reads were sampled at intervals of 250,000 reads, and the mean, de-duplicated depth of coverage was determined at each sampling level. (A, B, and C) Replicate captures were performed for each cardiology research panel. Eight different HapMap samples were prepared using KAPA HyperPrep Library Preparation Kit following the SeqCap EZ HyperCap Workflow User’s Guide v2.0. Captures were 8-plexed (using 125 ng of each sample library). Each capture was sequenced using an Illumina MiSeq sequencing instrument (MiSeq Reagent Kit v2; 2 x 101 bp). Data was subsampled to an average coverage depth of 250X.

Replicate 1 Replicate 2

Percent of reads on-target and percent of bases in padded target panel

Average of % mapped non-duplicate reads on-target (primary) Average of % bases in padded target (capture)

77.4

93.1

75.9

91.2

0

10

20

30

40

50

60

70

80

90

100

Replicate 1 Replicate 2

Fold 80 base penalty

Average of Fold 80 base penalty

3.0

2.5

1.5

2.0

1.0

0.5

0

1.47 1.45

Mean coverage vs reads

Replicate 1 Replicate 2

0

50

100

150

200

250

300

350

400

0 500000 1000000 1500000 2000000 2500000 3000000 3500000 4000000 4500000Replicate 1 Replicate 2

Percent of bases covered at 20X and 50X

Average of % bases ≥20 Average of % bases ≥50

100 99.98 100 99.97

0

20

40

60

80

100

120

A B

C D

Published by:

Roche Sequencing Solutions, Inc.4300 Hacienda DrivePleasanton, CA 94588

sequencing.roche.com

Data on file.For Research Use Only. Not for use in diagnostic procedures.HYPERCAP, KAPA and SEQCAP are trademarks of Roche. All other product names and trademarks are the property of their respective owners.© 2018 Roche Sequencing Solutions, Inc. All rights reserved. SEQ100225 05/18

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Design Share PortfolioSeqCap EZ Cardiology Panels are now available as part of Roche Sequencing Solutions’ Design Share Portfolio. Design Share makes it easy to access pre-designed NGS panels that are developed by Roche Sequencing Solutions or in collaboration with researchers around the world. Our newest panels include:

• SeqCap EZ Human Oncology Panel: Comprehensive pan-cancer coverage of 981 genes in a single research panel

• SeqCap EZ Inherited Disease Panel: 4,100 gene research panel on genes commonly associated with human genetic disorders

To access the full portfolio and design files, please visit: sequencing.roche.com/designshare.