| 1
TruSight
TM
Oncology 500
and TruSight
Oncology 500
High-Throughput
Enabling flexible, scalable
comprehensive genomic
profiling from FFPE samples
DATA SHEET
• Analyze multiple variant types and key biomarkers in
• Go from sample to results in 4–5 days using manual or
automated workflows that integrate library prep,
-
ondary analysis
• Generate accurate data and reliable results that meet
demanding performance specifications
• Keep samples in house and obtain data that is
relevant to the local institution and community
2 |
profiling has the potential to identify relevant genetic
16
A single, compre-
hensive assay to assess a wide range of biomarkers uses
less sample and returns results more quickly compared to
Oncology 500 and TruSight Oncology 500 High-Through-
put (Table 1
Analyze multiple tumor types and
biomarkers with a single workflow
TruSight Oncology 500 and TruSight Oncology 500
Figure 1Figure 2
content includes multiple variant types and key biomarkers
Table 2, , and Table 4-
nating the need to spend time and precious sample, such
Parameter TruSight Oncology 500 TruSight Oncology 500 High-Throughput
System
(research mode)
(research mode)
a
a
Sample throughput 8 samples per run
per run
run
Panel size
40 ng 40 ng 40 ng
40 ng
FFPE input requirement
Minimum recommendation of
2 mm
from FFPE tissue samples
Minimum recommendation of
2 mm
from FFPE tissue samples
Minimum recommendation of
2 mm
from FFPE tissue samples
Total assay time
variant report
variant report
variant report
Sequence run time 24 hr
Sequence run
Software version
5% VAF for small variants
fusions
amplifications
5% VAF for small variants
fusions (80 ng input)
amplifications
5% VAF for small variants
fusions (80 ng input)
amplifications
Analytical sensitivity
Analytical specificity
—TruSight Oncology 500 and TruSight Oncology 500 High-Throughput integrate into current lab
Prepare
samples
FFPE
specimens
Prepare libraries
and enrich
Sequence Call variants
Generate insights
and report
Commercial
DNA/RNA
extraction kits
Supports multiple
tissue types
TruSight Oncology 500
or
TruSight Oncology 500
High-Throughput
(automated
a
or manual)
DRAGEN TruSight
Oncology 500 Analysis
Software on local
DRAGEN server or
Connected Analytics
c
Multiple commercial options:
Connected Insights
d
Velsera CGW
NextSeq 550, NextSeq 550Dx,
b
NovaSeq 6000, or NovaSeq
6000Dx
b
Systems
NovaSeq X Series
Fusion
Splice
variant
Variant types detected by TruSight Oncology 500 and
oncology community to design TruSight Oncology 500
The resulting panels provide comprehensive coverage
of biomarkers commonly mutated in numerous cancer
types (
fusion and splice variants (Table 2,
significant coverage of key guidelines for multiple tumor
types (Figure 4
known correlations to responses,
9
and the tumor muta-
Table 4
10
-
ability of a comprehensive, streamlined workflow that
spans from sample input to final report (Figure 2
automated library preparation kits and methods, variant
calling tools, and interpretation and reporting software
enables a smooth workflow that can be completed in as
-
4 |
—TruSight Oncology 500 and TruSight Oncology 500
High-Throughput include key guideline biomarkers, emerging biomarkers, and pan-cancer biomarkers such as BRAF, NTRK1, NTRK2, NTRK3,
RET
*
Pan-cancer: BRAF, NTRK1, NTRK2, NTRK3, RET, MSI, TMB
Genes with biomarkers of clinical significance
*
Genes with
biomarkers
of potential
clinical
significance
†
Breast
BRCA1 BRCA2 ERBB2 ESR1 PALB2 PIK3CA 180
Colorectal
ERBB2 KRAS NRAS 166
Bone
EGFR ERG ETV1 ETV4 EWSR1 FEV FLI1 FUS H3F3A HEY1 IDH1
140
MDM2 NCOA2 SMARCB1
Lung
ALK EGFR ERBB2 KRAS MET NUTM1 ROS1 223
Melanoma
KIT NRAS ROS1 172
Ovarian
BRCA1 BRCA2 FOXL2 149
CNS
‡
APC ATRX CDKN2A CDKN2B EGFR H3F3A HIST1H3B HIST1H3C IDH1 IDH2 MYCN
140
PTCH1 RELA TERT TP53
Prostate
AR ATM BARD1 BRCA1 BRCA2 BRIP1 CDK12 CHEK1 CHEK2 FANCL FGFR2
151
FGFR3 PALB2 PTEN RAD51B RAD51C RAD51D RAD54L
Thyroid
HRAS KRAS NRAS TERT 165
Uterine &
cervical
BRCA2 EPC1 ERBB2 ESR1 FOXO1 GREB1 JAZF1 NCOA2 NCOA3 NUTM2A NUTM2B
138
PAX3 PAX7 PHF1 POLE SMARCA4 SUZ12 TP53 YWHAE
Other solid
tumors
ALK APC ARID1A ASPSCR1 ATF1 ATIC BAP1 BCOR BRCA1 BRCA2 CAMTA1
152
CARS CCNB3 CDK4 CDKN2A CIC CITED2 CLTC COL1A1 COL6A3 CREB1 CREB3L1
CREB3L2 CSF1 CTNNB1 DDIT3 DDX3X DNAJB1 DUX4 EED EGFR ERBB2 ERG
ETV1 ETV4 ETV6 EWSR1 FEV FGFR2 FGFR3 FLI1 FOXL2 FOXO1 FOXO4
FUS GLI1 HEY1 HGF HMGA2 IDH1 KRAS LEUTX MAML3 MDM2 MYB
MYOD1 NAB2 NCOA2 NF1 NFATC2 NFIB NR4A3 NRAS NUTM1 NUTM2A NUTM2B
PALB2 PATZ1 PAX3 PAX7 PDGFB PDGFRA PRKACA PRKD1 RANBP2 ROS1 SDHA
SDHB SDHC SDHD SMARCB1 SS18 SSX1 SSX2 SSX4 STAT6 SUZ12 TAF15
TCF12 TERT TFE3 TFEB TFG TP53 TPM3 TPM4 TRAF7 TSPAN31 VGLL2
WT1 WWTR1 YAP1 YWHAE ZC3H7B
| 5
ABL1 BCR CHEK1 EPHA7 FGF23 GSK3B IDH2 MAP3K1 NF2 PIK3CA RAD51D SMAD4 TGFBR2
ABL2 BIRC3 CHEK2 EPHB1 FGF3 H3F3A IFNGR1 MAP3K13 NFE2L2 PIK3CB RAD52 SMARCA4 TMEM127
ACVR1 BLM CIC ERBB2 FGF4 H3F3B INHBA MAP3K14 NFKBIA PIK3CD RAD54L SMARCB1 TMPRSS2
ACVR1B BMPR1A CREBBP ERBB3 FGF5 H3F3C INPP4A MAP3K4 NKX2-1 PIK3CG RAF1 SMARCD1 TNFAIP3
AKT1 BRAF CRKL ERBB4 FGF6 HGF INPP4B MAPK1 NKX3-1 PIK3R1 RANBP2 SMC1A TNFRSF14
AKT2
BRCA1
a
CRLF2 ERCC1 FGF7 HIST1H1C INSR MAPK3 NOTCH1 PIK3R2 RARA SMC3 TOP1
AKT3
BRCA2
a
CSF1R ERCC2 FGFR1 HIST1H2BD IRF2 MAX NOTCH2 PIK3R3 RASA1 SMO TOP2A
ALK BRD4 CSF3R ERCC3 FGFR2 HIST1H3A IRF4 MCL1 NOTCH3 PIM1 RB1 SNCAIP TP53
ALOX12B BRIP1 CSNK1A1 ERCC4 FGFR3 HIST1H3B IRS1 MDC1 NOTCH4 PLCG2 RBM10 SOCS1 TP63
ANKRD11 BTG1 CTCF ERCC5 FGFR4 HIST1H3C IRS2 MDM2 NPM1 PLK2 RECQL4 SOX10 TRAF2
ANKRD26 BTK CTLA4 ERG FH HIST1H3D JAK1 MDM4 NRAS PMAIP1 REL SOX17 TRAF7
APC C11orf30 CTNNA1 ERRFI1 FLCN HIST1H3E JAK2 MED12 NRG1 PMS1 RET SOX2 TSC1
AR CALR CTNNB1 ESR1 FLI1 HIST1H3F JAK3 MEF2B NSD1 PMS2 RFWD2 SOX9 TSC2
ARAF CARD11 CUL3 ETS1 FLT1 HIST1H3G JUN MEN1 NTRK1 PNRC1 RHEB SPEN TSHR
ARFRP1 CASP8 CUX1 ETV1 FLT3 HIST1H3H K AT6A MET NTRK2 POLD1 RHOA SPOP U2AF1
ARID1A CBFB CXCR4 ETV4 FLT4 HIST1H3I KDM5A MGA NTRK3 POLE RICTOR SPTA1 VEGFA
ARID1B CBL CYLD ETV5 FOXA1 HIST1H3J KDM5C MITF NUP93 PPARG RIT1 SRC VHL
ARID2 CCND1 DAXX ETV6 FOXL2 HIST2H3A KDM6A MLH1 NUTM1 PPM1D RNF43 SRSF2 VTCN1
ARID5B CCND2 DCUN1D1 EWSR1 FOXO1 HIST2H3C KDR MLL PAK1 PPP2R1A ROS1 STAG1 WISP3
ASXL1 CCND3 DDR2 EZH2 FOXP1 HIST2H3D KEAP1 MLLT3 PAK3 PPP2R2A RPS6KA4 STAG2 WT1
ASXL2 CCNE1 DDX41 FAM123B FRS2 HIST3H3 KEL MPL PAK7 PPP6C RPS6KB1 STAT3 XIAP
ATM CD274 DHX15 FAM175A FUBP1 HLA-A KIF5B MRE11A PALB2 PRDM1 RPS6KB2 STAT4 XPO1
ATR CD276 DICER1 FAM46C FYN HLA-B KIT MSH2 PARK2 PREX2 RPTOR STAT5A XRCC2
ATRX CD74 DIS3 FANCA GABRA6 HLA-C KLF4 MSH3 PARP1 PRKAR1A RUNX1 STAT5B YAP1
AURKA CD79A DNAJB1 FANCC GATA1 HNF1A KLHL6 MSH6 PAX3 PRKCI RUNX1T1 STK11 YES1
AURKB CD79B DNMT1 FANCD2 GATA2 HNRNPK KMT2B MST1 PAX5 PRKDC RYBP STK40 ZBTB2
AXIN1 CDC73 DNMT3A FANCE GATA3 HOXB13 KMT2C MST1R PAX7 PRSS8 SDHA SUFU ZBTB7A
AXIN2 CDH1 DNMT3B FANCF GATA4 IGF1 KMT2D MTOR PAX8 PTCH1 SDHAF2 SUZ12 ZFHX3
AXL CDK12 DOT1L FANCG GATA6 IGF1R KRAS MUTYH PBRM1 PTEN SDHB SYK ZNF217
B2M CDK4 E2F3 FANCI GEN1 IGF2 LAMP1 MYB PDCD1 PTPN11 SDHC TAF1 ZNF703
BAP1 CDK6 EED FANCL GID4 IKBKE LATS1 MYC PDCD1LG2 PTPRD SDHD TBX3 ZRSR2
BARD1 CDK8 EGFL7 FAS GLI1 IKZF1 LATS2 MYCL1 PDGFRA PTPRS SETBP1 TCEB1
BBC3 CDKN1A EGFR FAT1 GNA11 IL10 LMO1 MYCN PDGFRB PTPRT SETD2 TCF3
BCL10 CDKN1B EIF1AX FBXW7 GNA13 IL7R LRP1B MYD88 PDK1 QKI SF3B1 TCF7L2
BCL2 CDKN2A EIF4A2 FGF1 GNAQ INHA LYN MYOD1 PDPK1 RAB35 SH2B3 TERC
BCL2L1 CDKN2B EIF4E FGF8 GNAS HRAS LZTR1 NAB2 PGR RAC1 SH2D1A
TERT
b
BCL2L11 CDKN2C EML4 FGF9 GPR124 HSD3B1 MAGI2 NBN PHF6 RAD21 SHQ1 TET1
BCL2L2 CEBPA EP300 FGF10 GPS2 HSP90AA1 MALT1 NCOA3 PHOX2B RAD50 SLIT2 TET2
BCL6 CENPA EPCAM FGF14 GREM1 ICOSLG MAP2K1 NCOR1 PIK3C2B RAD51 SLX4 TFE3
BCOR CHD2 EPHA3 FGF19 GRIN2A ID3 MAP2K2 NEGR1 PIK3C2G RAD51B SMAD2 TFRC
BCORL1 CHD4 EPHA5 FGF2 GRM3 IDH1 MAP2K4 NF1 PIK3C3 RAD51C SMAD3 TGFBR1
BRCA1 and BRCA2
b. TERT
HIST2H3A, HIST2H3C, HLA-A, HLA-B, HLA-C, KMT2B, KMT2C, KMT2D, TERT
6 |
TruSight Oncology 500 High-Throughput panels
ABL1 EGFR FGFR2 MLL PAX3
AKT3 EML4 FGFR3 MLLT3 PAX7
ALK ERBB2 FGFR4 MSH2 PDGFRA
AR ERG FLI1 MYC PDGFRB
AXL ESR1 FLT1 NOTCH1 PIK3CA
BCL2 ETS1 FLT3 NOTCH2 PPARG
BRAF ETV1 JAK2 NOTCH3 RAF1
BRCA1 ETV4 KDR NRG1 RET
BRCA2 ETV5 KIF5B NTRK1 ROS1
CDK4 EWSR1 KIT NTRK2 RPS6KB1
CSF1R FGFR1 MET NTRK3 TMPRSS2
0 60 804020
100
Rectal cancer
Cervical cancer
Gastric cancer
Colon cancer
Neuroendocrine tumors
Breast cancer
Pancreatic adenocarcinoma
Non-small cell lung cancer
Uterine neoplasms
Melanoma
Percentage of genetic markers included
guidelines by cancer type—The graph provides examples of
Automate for efficiency
TruSight Oncology 500 and TruSight Oncology 500 High-
Throughput offer manual and automated options to
liquid-handling manufacturers, to produce fully automated
workflows for TruSight Oncology 500 assays that support
achieve the same high-quality results produced by manual
protocols, while reducing hands-on time by ~50%, enab-
Add tags for analytical specificity
During library preparation, unique molecular identifiers
11
Enrich libraries to focus efforts
to regions of interest, which are pulled down using
streptavidin-coated magnetic beads, and then eluted to
a useful strategy for analyzing specific genetic variants in
a given sample and reliably sequencing exomes or large
Hybrid–capture chemistry offers several advantages
over amplicon sequencing, including yielding data with
chemistry is fusion agnostic, enabling detection and
amplicon-based approaches, which require confirmatory
tests as false-positives can arise, the hybrid-capture
method is highly sensitive and can accurately characterize
Small variants Fusions
AKT1
ALK
BRAF
DDR2
EGFR
ERBB2
FGFR1
FGFR3
KRAS
MAP2K1
MET
NRAS
NTRK1
NTRK2
NTRK3
PIK3CA
PTEN
RET
TP53
Sequence 8–960 samples
TruSight Oncology 500 and TruSight Oncology 500
High-Throughput follow the same sample and library
*
Systems, which can
500 High-Throughput provides scalability to higher
6000 or
*
Systems, customers can batch from
This broad flexibility across platforms is enabled by the
availability of 192 unique indexes for TruSight Oncology
-
modate varying throughput levels (Table 5
index performs consistently to produce sequencing
Analyze data
Variant calling for TruSight Oncology 500 and TruSight
secondary analysis, either on premises using a local
Analytics, now with data streaming and autolaunch ca-
Scalable solution
Assay TruSight Oncology 500 TruSight Oncology 500 High-Throughput
System
(research mode)
System (research mode)
a
Flow cell High-output SP S1 S2 S4
8 16 192 192 480
8 |
proprietary algorithms that remove errors, artifacts, and
germline variants, resulting in highly accurate variant
calling performance with an analytical specificity of
-
eficial when it is critical to know the exact number of
†
500 show concordant results (,
Table 6
read the Security, privacy, and compliance with
Variant insights and report generation are available
potentially thousands of variants, biologically relevant
variants can be filtered and prioritized into a final, custom-
Oncology 500 Analysis Software
biopsy
samples
Average time for analysis to complete
a
b
c
8 2 hr
16 12 hr
18 hr 5 hr
24 hr 10 hr
† Previous generation of TruSight Oncology 500 software (not
Proven, reliable results
Although TruSight Oncology 500 and TruSight Oncology
500 High-Throughput were designed to run on separate
sequencing platforms with different throughput options,
the assays have the same genomic content and per-
TruSight Oncology 500 and TruSight Oncology 500
methods deliver a qualitative result describing samples
-
ment with the TruSight Oncology 500 assays interro-
Figure 5
12
TruSight Oncology 500 panels combine comprehensive
genomic content with sophisticated informatics algorithms
-
dant with whole-exome studies (Figure 6,
12
The
11
-
amination, oxidation) enables analytical sensitivity as low
Metric Value
Percent positive agreement
Percent negative agreement
Overall percent agreement
| 9
0%
TruSight Oncology 500
High-Throughput (DRAGEN App)
percent unstable sites
TruSight Oncology 500
High-Throughput percent unstable
sites
TruSight Oncology 500
High-Throughput (Local App) percent
unstable sites
R
2
= 0.999
60%
20%
40%
0%
20% 40% 60% 80%
80%
TruSight Oncology 500
percent unstable sites
TruSight Oncology 500
percent unstable sites
MSI-PCR
percent unstable sites
0% 20% 40% 60% 80%
60%
20%
40%
0%
80%
60%
20%
40%
0%
High Stable
MSI Status:
MSI-H MSS
MSI Status:
MSI-H MSS
R
2
= 0.990
A.
B. C.
A.
B. C.
TruSight Oncology 500
High-Throughput
NovaSeq mutations/Mb
R
2
= 0.998
R
2
= 0.931
TruSight Oncology 500
NextSeq mutations/Mb
TruSight Oncology 500
mutations/Mb
WES mutations/Mb
150
100
50
0
60
40
20
0
0 50 100 1500 20 40 150
80
10 |
-
ciated genes and can call amplifications with a limit of
change (, Table 8
Detection of BRCA large rearrangements
A BRCA
TruSight Oncology 500 analysis workflow enables exon-
BRCA
Table 9
Highly sensitive variant detection from FFPE
samples
One benefit of target enrichment chemistry is the use
of probes designed large enough to impart high binding
specificity, but also allow hybridization to targets con-
dropouts in the presence of both natural allelic variations
and sequence artifacts introduced from FFPE tissue
Figure 8, Table 10
Mean fold change
Gene
Oncology 500 v2
TruSight Oncology 500
AR
BRAF
BRCA1
BRCA2
CCND1
CCNE1
CDK4
CDK6
CHEK2
EGFR
ERBB2
FGF10
FGF19
FGFR1
KRAS
MDM2
MDM4
MET
MYC
MYCN
The information in this table shows examples of high concordance of data analyzed
level changes, and gene fusions that are undetectable
14,15
The TruSight Oncology
500 assays detect and characterize fusions agnostic
is recommended for use with TruSight Oncology 500
to detect variants expressed at mid to high levels with
fusions present at low concentrations (Table 11
| 11
BRCA
BRCA 1/2
BRCA1 loss exon 8
BRCA2 loss exon 21–24
BRCA1 loss exon 14–24
BRCA1 loss exon 21–24
BRCA1
BRCA1
BRCA2
BRCA1
BRCA1 gain exon 5–16
BRCA1
BRCA1 gain exon 1–16
BRCA1
BRCA2 gain exon 25
BRCA2
BRCA2
BRCA1 loss exon 22
Gene Mutation
Oncology 500 v2
AKT1 5%
APC 8%
BRAF V600E
CTNNB1 T41A 8%
EGFR
EGFR
FGFR3 6%
FOXL2
GNAS
IDH1
KIT D816V 8%
KRAS G12D
NOTCH1 P668S 5%
NRAS
PIK3CA E545G 5%
RET M918T 8%
TP53
Variant type: complex variant
EGFR
Variant type: insertion
APC
ERBB2
Variant type: deletion
FBXW7 5%
PTEN
TP53 6%
The information in this table shows examples of concordance between data analyzed
12 |
amount
Tissue
40
ng
60
ng
80
ng
ALK-EML4 15 21 40
EGFR-RAB3IP 5 9 19
EGFR-METTL1 25 84
BRCA1-MPP2 25 28 29
ALK-BRE 112 128 Sarcoma
CCDC170-ESR1 122 59 168 Kidney
MYC-MRPL13 52
MYC-STK3 11 28
ROS1;GOPC-ENC1
ROS1;GOPC-CD74 104 92 141
ANKUB1; RNF13-ETV5;DGKG 29 45
NTRK3-SEMA6A 16 25 Skin
RET-NCOA4 154 Thyroid
EWSR1-ATF1 19 Sarcoma
EWSR1-CBY1 44 Sarcoma
BRCA2-NRXN3 60 84
FLT3-SMOX 50 54
FLT3-VWA8 29 51 69
FLT3-LCP1 12
Splice variant
ARv7 26 46
EGFR v3 884
EGFR v3 1249 1614 2049
Plan for the future
TruSight Oncology 500 and TruSight Oncology 500
High-Throughout integrate easily into labs currently using
multiple independent, single biomarker assays into one
assay, labs can save sample, time, and money, while
Enhanced product attributes
efficiency, TruSight Oncology 500 products
‡
feature:
• Advanced change notification‒-
oratories six months before any significant changes
are made to a product in the TruSight Oncology 500
portfolio
†
• Certificate of Analysis‒Every TruSight Oncology 500
product
†
-
tains the product has met its predetermined product
release specifications and quality
• Extended shelf life‒The minimum guaranteed shelf life
for TruSight Oncology 500 reagents is extended to six
months, reducing the risk of product expiration and en-
abling labs to use reagents according to current testing
needs
Summary
TruSight Oncology 500 and TruSight Oncology 500
present in guidelines and clinical trials, in a single assay
hybrid-capture with sophisticated informatics reduces
errors and yields high-quality data, even from FFPE
labs can increase their batch sizes and process more
samples per week across a broad range of sequencing
analysis enables TruSight Oncology 500 to improve lab
Ordering information: TruSight Oncology 500
Sample
type
Velsera
included
Product
Manual
a
indexes, 48 samples)
Velsera
a
b
20028214
b
a
20028215
plus Velsera
a
b
20028216
b
Automated
a
Kit (16 indexes, 64 samples)
20045504
Kit, plus Velsera
a
20045506
b
64 samples)
20045505
b
a
20045508
Automation Kit, plus Velsera
a
20045509
b
20045990
Velsera
b
20045991
TruSight Oncology 500 and TruSight Oncology
500 High-Throughput
analysis
Analytics
14 |
Ordering information: TruSight Oncology 500 and TruSight Oncology 500 High-Throughput
Sample
type
Velsera
included
Automation
Product Product
Manual
Throughput Kit
a
sales
Throughput Kit, with Velsera
a
sales
Throughput Kit
a
Throughput, with Velsera
a
Throughput Kit
a
Throughput Kit, with Velsera
a
Throughput Kit
a
Throughput Kit, with Velsera
a
Automated
Throughput Automation Kit
a
Throughput Automation Kit
a
plus Velsera
Throughput Automation Kit
a
20049285
Throughput Automation Kit
a
plus Velsera
Throughput Automation Kit
a
20049282
Throughput Automation Kit
a
plus Velsera
Throughput Automation Kit
a
20049284
Throughput Automation Kit
a
plus Velsera
| 15
Ordering information: TruSight Oncology 500
High-Throughput, continued
Ordering information: Analysis options
On-premises variant calling
Product Product
20040619
Manual
Automated
96 samples)
20066404
96 samples)
Subscription
Subscription
20049422
Analytics, 1 credit
Analytics Starter Pack, 1000 credits
Analytics, 5000 credits
20042040
20085595
Analytics, 50,000 credits
20042041
Analytics, 100,000 credits
20042042
| 16
The land-
scape of kinase fusions in cancerNat Commun
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programOncotarget
High-Throughput Genom-
Cancer Discov
Multicenter Fea-
-
JAMA Oncol
JAMA Oncol.
Mutational landscape of
metastatic cancer revealed from prospective clinical se-
quencing of 10,000 patientsNat Med
-
Cancer Immunol Res
Tumor mutational
load predicts survival after immunotherapy across multiple
cancer typesNat Genet
-
fda-approves-first-line-immunotherapy-patients-msi-hdmmr-
metastatic-colorectal-cancer
-
ab-adults-and-children-tmb-h-solid-tumors
The landscape of
somatic copy-number alteration across human cancersNature
-
Nat Commun
Am J Hum Genet
