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GUIDES — RESEARCH DESK

From swab to sequencer

A plain-spoken map of hereditary-cancer testing: how a saliva tube or buccal swab becomes sequenced data, how laboratories classify what they find, and how a report finally reaches a clinic. Educational explainers, citation-minded, never a sales pitch.

~13%
of women in the general population develop breast cancer in a lifetime
NCI
>60%
lifetime breast-cancer risk in BRCA1/BRCA2 carriers
NCI
5 tiers
ACMG/AMP variant classes, from Pathogenic to Benign
ACMG/AMP 2015
28
weighted evidence criteria behind a classification call
ACMG/AMP 2015

The end-to-end workflow

Every hereditary-cancer result travels the same path. Understanding each handoff makes a report far less opaque — and explains why turnaround takes weeks, not minutes.

  1. 01

    1 · Indication & counselling

    A personal or family history flags possible hereditary risk. A clinician or genetic counsellor frames what a test can and cannot answer before any sample is taken.

  2. 02

    2 · Collection

    DNA is captured from saliva (OG-500), a buccal swab, or a blood draw (EDTA vacutainer). The choice trades sample yield against convenience and stability.

  3. 03

    3 · Stabilise & ship

    Lysis or preservation buffer protects the DNA; the tube is triple-packed as UN3373 Category B and shipped at ambient temperature under packing instruction P650.

  4. 04

    4 · Extraction

    The lab isolates genomic DNA, checks yield and purity (A260/A280), and normalises concentration before library preparation.

  5. 05

    5 · Sequencing

    A targeted multi-gene panel is enriched and read on a flow cell — typically ~100× depth for germline panels, against the ~30× standard for whole-genome sequencing.

  6. 06

    6 · Variant calling & classification

    Reads are aligned, variants called, then each is graded against the ACMG/AMP framework: Pathogenic, Likely pathogenic, VUS, Likely benign, or Benign.

  7. 07

    7 · Report & return

    A clinical report goes back to the ordering clinician, who interprets findings in context — risk figures, surveillance options, and onward referral.

Collection methods at a glance

The first decision in any workflow is how to capture DNA. Yields and stability below reflect manufacturer technical specifications and common laboratory practice; treat them as typical ranges, not guarantees.

MethodTypical sampleStabilityNotes
Saliva — OG-500~2 mL saliva + bufferYears at room temperatureHigh total yield; lysis buffer stabilises DNA immediately on capping.
Buccal swabCheek-cell scrapeWeeks, ambient (dry/buffered)Low burden, child-friendly; lower yield than saliva.
Blood draw — EDTA vacutainer2–10 mL whole bloodDays refrigerated; longer if processedHighest-quality DNA; needs phlebotomy and tighter handling.

Table 1. Common germline DNA collection methods and their handling profile.

Sequencing depth by approach

"Depth" is how many times each base is read. Higher depth means more confident variant calls — targeted hereditary-cancer panels are read far deeper than a whole genome to maximise sensitivity over a focused set of genes.

Whole-genome (germline standard)30 × depth

≥30× gold standard for SNV/indel discovery

Whole-exome (germline)100 × depth

≥50× recommended, ~100× preferred

Targeted multi-gene panel100 × depth

Deep coverage over a curated gene set

swab → sequencer
CLASSIFICATION

What "Pathogenic" — and "uncertain" — really mean

A sequenced variant is not a verdict on its own. Under the 2015 ACMG/AMP framework, laboratories weigh up to 28 criteria — population frequency, computational prediction, functional and segregation data — to place each variant in one of five tiers: Pathogenic, Likely pathogenic, Variant of Uncertain Significance (VUS), Likely benign, or Benign. The "likely" tiers carry a target confidence of about 90%.

A VUS is the honest middle: real, but not yet understood. It is not a diagnosis, and classifications can be revised as evidence accumulates — one reason the original PROMPT registry mattered. Personal decisions belong with a clinician or genetic counsellor, not a label alone.

A registry's value is the boring part: tracking the same variant across many people and many years, so a result classed "uncertain" today can be resolved tomorrow.
PROMPT Registry · research desk

Begin at the beginning

Follow the pathway in order, or jump to the hub that answers your question. Everything here is an educational explainer — for a personal decision, speak with a clinician or genetic counsellor.