1. TS Tumor Panel (15 Genes) Overview
Technical training
TS Tumor Panel (15 Genes) Overview
Targets relevant regions in 15 cancer-associated genes
Content created by NCCN, ESMO, KOLs, and leading pharma companies
Enhanced Sample-to-Data
Detailed QC steps and simplified pre-defined variant report
Workflow optimized for Oncology customers
Low FFPE input (20ng) and quick TAT (~36 hours)
Sensitive, High-Confidence Variant Detection
Identifies somatic mutations at 5% variant allele frequency with >500x coverage
Gastric

2. TruSight Tumor Panel (15 Genes) Sample-To-Data Workflow Solution
Technical training
TruSight Tumor Panel (15 Genes) Sample-To-Data Workflow Solution
FFPE specimens
Sample
Library Prep
Sequencing
Analysis
Tissue Guidelines
Extraction Methods DNA Quant Guidelines
FFPE optimized
Low DNA-input
Multiplex PCR
Library QC Guidelines
MiSeq V3
Leading Sequencers
Sequencing QC
MSR
Pre-Defined Report
Variant Studio
3rd Party Annotation
Text underlined is additional QC/analysis that is included in TST 15. These are things that were not included for previous kits.
Extends beyond library preparation to enable a more complete solution
For Research Use Only. Not for use in diagnostic procedures.

3. TruSight Tumor Panel (15 Genes) Sample-To-Data Turn Around Time
Technical training
TruSight Tumor Panel (15 Genes) Sample-To-Data Turn Around Time
DAY 1
DAY 2
DAY 3
LIBRARY PREP
7 hours
SEQUENCING
27 hours
ANALYSIS
1 hour
DNA to Analysis in less than 3 days

4. TruSight Tumor Panel (15 Genes) Content List
Key Benefits
Gene content: created with industry guidance (NCCN, CAP), leading KOLs, and pharma
Relevance: Selected genes have the most analytic relevance for translational tumor research
Focused: Small panel ensures quick and comprehensive sample-to-data turnaround time
Gene
Associated Cancers
Target
AKT1
Breast
E17K
BRAF
Melanoma*, Colon* Lung
VK600E/K/R/M
EGFR
Lung*
Exon 19, 20, 21(L858R), G719A/X, S7681, T790M, L861Q
ERBB2
Breast, Lung
A775_G776insYVMA
FOXL2
Ovary
C134W
GNA11
Melanoma
Q209L
GNAQ
KIT
Gastric, Melanoma*
Exons 8, 9, 10, 11, 13, 14, 17, 18
KRAS
Colon*, Gastric, Lung*
Codons 12, 13, 59, 61, 117, 146
MET
Lung*, Colon, Gastric
Focal Amplification
NRAS
Colon*
PDGFRA
Gastric, Melanoma
Exons 12, 14, 18
PIK3CA
Lung, Breast, Prostate
Exons 9, 20
RET
M918T
TP53
Lung, Melanoma, Ovary, Colon
Full CDS
*Indicates in NCCN guideline for cancer type

5. TruSight Tumor Panel (15 Genes) Recommended QC
Technical training
TruSight Tumor Panel (15 Genes) Recommended QC
STEP
QC Method
QC METRIC
FFPE Tissue
Sectioning of tissue with microtome
≥ 140 mm2 (non-melanoma)
≥ 20% tumor
DNA Extraction
Input concentration fluorometry
≥ 2 ng/µL
Library Prep
Final concentration by fluorometry
≥ 20 ng/µL final yield
Final fragment size on gel (except NTC)
~350bp
Sequencing
Review of analysis report
>93.5% bases >500X

6. TS Tumor Panel (15 Genes) Library Prep
Technical training
TS Tumor Panel (15 Genes) Library Prep
Mix A
Mix B
5 Steps!
Multiplex PCR
Index PCR
Clean Up
Validate
Sequence

7. TS Tumor Panel (15 Genes) Multiplex PCR Chemistry
Technical training
TS Tumor Panel (15 Genes) Multiplex PCR Chemistry
Two libraries per sample: Pool A & Pool B

8. TruSight Tumor Panel (15 Genes) Analysis and Reporting
Technical training
TruSight Tumor Panel (15 Genes) Analysis and Reporting
Illumina Experiment Manager v1.10
Adds TruSight Tumor Panel (15 Genes) workflow and library prep
IEM v1.10 will be released upon TST 15 FCS
MiSeq Software v2.6
MSR TruSight Tumor Panel (15 Genes) Workflow
Alignment and variant calling for TST 15.
Creates pre-defined filtered report for easy data interpretation.
MCS/MSR 2.6 will be released upon TST 15 FCS
VariantStudio
TruSight Tumor 15 customers receive license for VariantStudio
Enables customized filtering of variants and reporting if pre-defined report is not sufficient

9. Technical training
TruSight Tumor Panel (15 Genes) Pre-defined report makes variant calling easy
Streamlined variant filtering and classification
Easy to use automated analysis

10. TruSight Tumor Panel (15 Genes) Data Specifications
Technical training
TruSight Tumor Panel (15 Genes) Data Specifications
PARAMETER
DETAIL
DNA input requirements
20 ng
Panel size
44 kb
Amplicon size
~150–175 bp
Throughput
8 Samples per MiSeq v3 run
Variant frequency 5%
Coverage
> 93.5% bases covered at > 500×

11. TruSight Tumor Panel (15 Genes) FFPE input
Technical training
TruSight Tumor Panel (15 Genes) FFPE input
Library concentration vs. ΔCq. ΔCq is a metric used to assess the quality of FFPE DNA. Higher values reflect poorer quality DNA. A library yield of 20ng/ul is recommended for TruSight Tumor Panel (15 Genes).

12. TruSight Tumor Panel (15 Genes) TST 15 vs TST 26 variant calling
Technical training
TruSight Tumor Panel (15 Genes) TST 15 vs TST 26 variant calling
Gene
Coordinate
Variant
Protein
TST 26
TST 15 Smp1
Rep1
MixA
Rep2
MixB
PDGFRA
A>G/G
567P
99.6
99.84
99.82
99.73
99.54
EGFR,EGFR-AS1
G>A/A
99.64
99.93
99.98
TP53
C>C/A
154G/V
14.4
18.6
20.33
G>C/C
72P/R
99.52
99.65
99.76
99.77
99.74
100% of variants detected by TruSight Tumor (26 Genes) were detected by TruSight Tumor (15 Genes) and reported at similar frequencies.

13. TST (15 Genes) – Kit Configuration
Technical training
TST (15 Genes) – Kit Configuration
Library Prep Kit Content
TruSight Tumor Kit (15 Genes)
TruSight Tumor Panel (15 Genes)
24 Samples, 48 libraries
3 Boxes
Includes Library Prep reagents, 24 index combinations, and purification beads
Cat # OP
Includes 1 Library Prep AND
3 MiSeq Sequencing Kits
Cat# OP
1 Library Prep Kit ONLY