INVITAE DIAGNOSTIC TESTING RESULTS

Patient name: Ester Perez Poblete Sample type: Blood Report date: 14-SEP-2022
DOB: 26-AUG-1996 Sample collection date: 01-SEP-2022 Invitae #: RQ4035649
Sex assigned at birth: Female Sample accession date: 06-SEP-2022 Clinical team: Manuel Álvarez
Gender:
Patient ID (MRN): 19191276-4

Reason for testing Test performed

Diagnostic test for a personal history of disease Sequence analysis and deletion/duplication testing of the 26 genes listed
in the Genes Analyzed section.
Invitae Breast Cancer Panel
Add-on Preliminary-evidence Genes for Breast Cancer

RESULT: NEGATIVE

About this test

This diagnostic test evaluates 26 gene(s) for variants (genetic changes) that are associated with genetic disorders. Diagnostic genetic
testing, when combined with family history and other medical results, may provide information to clarify individual risk, support a
clinical diagnosis, and assist with the development of a personalized treatment and management strategy.

Next steps

This test did not identify any pathogenic variants known to cause disease. This result should be discussed with a healthcare
provider, such as a genetic counselor, to learn about the appropriate next steps for further evaluation. Clinical follow up may still
be warranted. This result should be interpreted within the context of additional laboratory results, family history and clinical
findings.

Register your test at [Link]/patients to download a digital copy of your results. You can also access educational
resources about how your results can help inform your health.

Laboratory Director: Neng Chen, PhD, FACMG, NYCQ, CGMB

Invitae 1400 16th Street, San Francisco, CA 94103 · clientservices@[Link] · 800.436.3037
© 2022 Invitae Corporation Page 1 of 5
 INVITAE DIAGNOSTIC TESTING RESULTS
Patient name: Ester Perez Poblete DOB: 26-AUG-1996
Invitae #: RQ4035649

Clinical summary

No reportable genetic variants were identified by this analysis, however, this individual may still be at risk for certain medical
conditions based on other factors such as family history, genetic causes not evaluated with this test, or other environmental
influences. Follow up of this individual and surveillance of their family members may still be indicated.

Laboratory Director: Neng Chen, PhD, FACMG, NYCQ, CGMB

Invitae 1400 16th Street, San Francisco, CA 94103 · clientservices@[Link] · 800.436.3037
© 2022 Invitae Corporation Page 2 of 5
 INVITAE DIAGNOSTIC TESTING RESULTS
Patient name: Ester Perez Poblete DOB: 26-AUG-1996
Invitae #: RQ4035649

Genes analyzed

This table represents a complete list of genes analyzed for this individual, including the relevant gene transcript(s). If more than one transcript is listed for a single
gene, variants were reported using the first transcript listed unless otherwise indicated in the report. An asterisk (*) indicates that this gene has a limitation. Please
see the Limitations section for details. Results are negative unless otherwise indicated in the report. Benign and Likely Benign variants are not included in this
report and in specific scenarios variants of uncertain significance in the requisitioned gene(s) may not be included in this report. These variants are available upon
request.

GENE TRANSCRIPT
ABRAXAS1 NM_139076.2
AKT1 NM_005163.2
ATM* NM_000051.3
BARD1 NM_000465.3
BRCA1 NM_007294.3
BRCA2 NM_000059.3
BRIP1 NM_032043.2
CDH1 NM_004360.3
CHEK2 NM_007194.3
FANCC NM_000136.2
FANCM NM_020937.2
MRE11 NM_005591.3
MUTYH NM_001128425.1
NF1* NM_000267.3
PALB2 NM_024675.3
PIK3CA NM_006218.2
PTEN* NM_000314.4
RAD51C NM_058216.2
RAD51D NM_002878.3
RECQL* NM_002907.3
RINT1 NM_021930.4
SDHB NM_003000.2
SDHD NM_003002.3
STK11 NM_000455.4
TP53 NM_000546.5
XRCC2 NM_005431.1

Laboratory Director: Neng Chen, PhD, FACMG, NYCQ, CGMB

Invitae 1400 16th Street, San Francisco, CA 94103 · clientservices@[Link] · 800.436.3037
© 2022 Invitae Corporation Page 3 of 5
 INVITAE DIAGNOSTIC TESTING RESULTS
Patient name: Ester Perez Poblete DOB: 26-AUG-1996
Invitae #: RQ4035649

Methods

Genomic DNA obtained from the submitted sample is enriched for targeted regions using a hybridization-based protocol, and sequenced using
Illumina technology. Unless otherwise indicated, all targeted regions are sequenced with ≥50x depth or are supplemented with additional analysis.
Reads are aligned to a reference sequence (GRCh37), and sequence changes are identified and interpreted in the context of a single clinically
relevant transcript, indicated below. Enrichment and analysis focus on the coding sequence of the indicated transcripts, 20bp of flanking intronic
sequence, and other specific genomic regions demonstrated to be causative of disease at the time of assay design. Promoters, untranslated
regions, and other non-coding regions are not otherwise interrogated. For some genes only targeted loci are analyzed (indicated in the table
above). Exonic deletions and duplications are called using an in-house algorithm that determines copy number at each target by comparing the
read depth for each target in the proband sequence with both mean read-depth and read-depth distribution, obtained from a set of clinical
samples. Markers across the X and Y chromosomes are analyzed for quality control purposes and may detect deviations from the expected sex
chromosome complement. Such deviations may be included in the report in accordance with internal guidelines. Confirmation of the presence
and location of reportable variants is performed based on stringent criteria established by Invitae (1400 16th Street, San Francisco, CA 94103,
#05D2040778), as needed, using one of several validated orthogonal approaches (PubMed ID 30610921). The following analyses are performed if
relevant to the requisition. For PMS2 exons 12-15, the reference genome has been modified to force all sequence reads derived from PMS2 and
the PMS2CL pseudogene to align to PMS2, and variant calling algorithms are modified to support an expectation of 4 alleles. If a rare SNP or
indel variant is identified by this method, both PMS2 and the PMS2CL pseudogene are amplified by long-range PCR and the location of the variant
is determined by Pacific Biosciences (PacBio) SMRT sequencing of the relevant exon in both long-range amplicons. If a CNV is identified, MLPA
or MLPA-seq is run to confirm the variant. If confirmed, both PMS2 and PMS2CL are amplified by long-range PCR, and the identity of the fixed
differences between PMS2 and PMS2CL are sequenced by PacBio from the long-range amplicon to disambiguate the location of the CNV.
Technical component of confirmatory sequencing is performed by Invitae Corporation (1400 16th Street, San Francisco, CA 94103,
#05D2040778). For C9orf72 repeat expansion testing, hexanucleotide repeat units are detected by repeat-primed PCR (RP-PCR) with fluorescently
labeled primers followed by capillary electrophoresis. Interpretation Reference Ranges: Benign (Normal Range): <25 repeat units, Uncertain: 25-30
repeat units, Pathogenic (Full Mutation): >=31 repeat units. A second round of RP-PCR utilizing a non-overlapping set of primers is used to
confirm the initial call in the case of suspected allele sizes of 22 or more repeats. For RNA analysis of the genes indicated in the Genes Analyzed
table, complementary DNA is synthesized by reverse transcription from RNA derived from a blood specimen and enriched for specific gene
sequences using capture hybridization. After high-throughput sequencing using Illumina technology, the output reads are aligned to a reference
sequence (genome build GRCh37; custom derivative of the RefSeq transcriptome) to identify the locations of exon junctions through the detection
of split reads. The relative usage of exon junctions in a test specimen is assessed quantitatively and compared to the usage seen in control
specimens. Abnormal exon junction usage is evaluated as evidence in the Sherloc variant interpretation framework. If an abnormal splicing
pattern is predicted based on a DNA variant outside the typical reportable range, as described above, the presence of the variant is confirmed by
targeted DNA sequencing. RNA sequencing is performed by Invitae Corporation (1400 16th Street, San Francisco, CA 94103, #05D2094793).
Technical component of Fibroblast cell-culturing and gDNA extraction from skin punch biopsy is performed by Invitae Corporation (5 Technology
Drive, Irvine CA 92618, #05D1052995).
A PMID is a unique identifier referring to a published, scientific paper. Search by PMID at [Link]
An rsID is a unique identifier referring to a single genomic position, and is used to associate population frequency information with sequence
changes at that position. Reported population frequencies are derived from a number of public sites that aggregate data from large-scale
population sequencing projects, including ExAC ([Link] gnomAD ([Link] and dbSNP
([Link]
A MedGen ID is a unique identifier referring to an article in MedGen, NCBI's centralized database of information about genetic disorders and
phenotypes. Search by MedGen ID at [Link] An OMIM number is a unique identifier referring to a
comprehensive entry in Online Mendelian Inheritance in Man (OMIM). Search by OMIM number at [Link]
Invitae uses information from individuals undergoing testing to inform variant interpretation. If "Invitae" is cited as a reference in the variant
details this may refer to the individual in this requisition and/or historical internal observations.

Limitations

Based on validation study results, this assay achieves >99% analytical sensitivity and specificity for single nucleotide variants, insertions and deletions
<15bp in length, and exon-level deletions and duplications. Invitae's methods also detect insertions and deletions larger than 15bp but smaller than a full

Laboratory Director: Neng Chen, PhD, FACMG, NYCQ, CGMB

Invitae 1400 16th Street, San Francisco, CA 94103 · clientservices@[Link] · 800.436.3037
© 2022 Invitae Corporation Page 4 of 5
 INVITAE DIAGNOSTIC TESTING RESULTS
Patient name: Ester Perez Poblete DOB: 26-AUG-1996
Invitae #: RQ4035649

exon but sensitivity for these may be marginally reduced. Invitae’s deletion/duplication analysis determines copy number at a single exon resolution at
virtually all targeted exons. However, in rare situations, single-exon copy number events may not be analyzed due to inherent sequence properties or
isolated reduction in data quality. Certain types of variants, such as structural rearrangements (e.g. inversions, gene conversion events, translocations,
etc.) or variants embedded in sequence with complex architecture (e.g. short tandem repeats or segmental duplications), may not be detected.
Additionally, it may not be possible to fully resolve certain details about variants, such as mosaicism, phasing, or mapping ambiguity. Unless explicitly
guaranteed, sequence changes in the promoter, non-coding exons, and other non-coding regions are not covered by this assay. Please consult the test
definition on our website for details regarding regions or types of variants that are covered or excluded for this test. This report reflects the analysis of an
extracted genomic DNA sample. While this test is intended to reflect the analysis of extracted genomic DNA from a referred patient, in very rare cases the
analyzed DNA may not represent that individual’s constitutional genome, such as in the case of a circulating hematolymphoid neoplasm, bone marrow
transplant, blood transfusion, chimerism, culture artifact or maternal cell contamination. Invitae's RNA analysis is not designed for use as a stand-alone
diagnostic method and cannot determine absolute RNA levels. Results from the RNA analysis may not be informative for interpreting copy number gains.
ATM: Sequencing analysis for exons 6, 24, 43 includes only cds +/- 10 bp. RECQL: Sequencing analysis for exons 3, 5, 15 includes only cds +/- 10 bp. NF1:
Sequencing analysis for exons 2, 7, 25, 41, 48 includes only cds +/- 10 bp. PTEN: Sequencing analysis for exons 8 includes only cds +/- 10 bp.

Disclaimer

DNA studies do not constitute a definitive test for the selected condition(s) in all individuals. It should be realized that there are possible sources of error.
Errors can result from trace contamination, rare technical errors, rare genetic variants that interfere with analysis, recent scientific developments, and
alternative classification systems. This test should be one of many aspects used by the healthcare provider to help with a diagnosis and treatment plan,
but it is not a diagnosis itself. This test was developed and its performance characteristics determined by Invitae. It has not been cleared or approved by
the FDA. The laboratory is regulated under the Clinical Laboratory Improvement Act (CLIA) as qualified to perform high-complexity clinical tests (CLIA ID:
05D2040778). This test is used for clinical purposes. It should not be regarded as investigational or for research.

This report has been reviewed and approved by:

Matteo Vatta, Ph.D., FACMG

Clinical Molecular Geneticist

Laboratory Director: Neng Chen, PhD, FACMG, NYCQ, CGMB

Invitae 1400 16th Street, San Francisco, CA 94103 · clientservices@[Link] · 800.436.3037
© 2022 Invitae Corporation Page 5 of 5
 NEGATIVE RESULTS GUIDE

What negative results mean for you

Your genetic test results were negative. This means that no significant genetic changes (“pathogenic variants” or
“mutations”) were found. Your risk for disease could still be influenced by a combination of unidentified genetic,
personal, lifestyle and/or environmental risk factors.

Create a plan with your healthcare provider

Whether or not you develop a disease is not determined by your genetics alone. It is still important to share your
genetic test results with your healthcare provider so they can help you make informed medical decisions.

What negative results mean for your family

Your genetic test was negative, however, your family members have their own unique genetic makeup. Genetic
testing can help them understand their overall chance of developing a genetic disease.

We (and others) are here to help

Although your test didn’t find any genetic changes, you may still have questions about your results or your personal
or family medical history. A genetic counselor can help.

Log in to your patient portal ([Link]) to view your results, search for a local or Invitae genetic counselor, or
join Invitae’s Patient Insight Network (PIN), a community where you can connect with other patients and share your
experience.

This information in this results guide is meant to be used along with your genetic test results and other health information. It is not meant to replace a discussion with your
healthcare provider and should not be considered or interpreted as medical advice.

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