SNaPshot multiplex PCR
Overview
SNaPshot (Applied Biosystems) is a dideoxy single-base-extension assay run after multiplex PCR amplification. Primers are extended by a single fluorescently labelled dideoxynucleotide complementary to the target position; size and colour of the extension product identify the base at each interrogated locus. The method is well-suited to focused hotspot genotyping panels covering tens of positions with high sensitivity in FFPE or frozen tissue.
Used by
- PMID:30325352 — SNaPshot multiplex PCR used to detect single-nucleotide mutations in EGFR (exons 18, 19, 20, 21) and KRAS (exon 2, codons 12 and 13) in 206 and 205 of 211 NSCLC subjects, respectively, in the nsclc-radiogenomics-stanford radiogenomic dataset; ALK/EML4 fusion status was assessed separately by FISH PMID:30325352.
Notes
- SNaPshot is a clinical-grade targeted assay; it detects only the specific codons included in the primer panel and cannot discover novel hotspots or indels outside the targeted regions.
- In the radiogenomics context, SNaPshot results are used as ground-truth mutation labels for training/validating CT-based radiogenomic prediction models PMID:30325352.
Sources
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