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Unknown,Transcriptomics,Genomics,Proteomics

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Effect of a KRAS peptide nucleic acid clamp on the Ion Torrent sequencing protocol

ABSTRACT: To test whether the addition of a peptide nucleic acid (PNA) clamp, which binds WT KRAS at codon 12, can increase the efficacy of mutation detection for KRASG12D within a targeted NGS setting. We tested the effect of clamping the wild-type KRAS sequence in a reference standard (Tru-Q 7, 1.3% Tier from Horizon Diagnostics, Cambridge, UK) with a KRAS c.35G>A mutation (KRASG12D) at an allelic frequency (AF) of 1.3% assessed by digital droplet PCR (ddPCR). We then re-tested the PNA on circulating-free DNA from a patient harbouring a KRASG12D mutation (at an AF of 3.2%, determined by ddPCR). Multiple runs were conducted using 10, 5, 2.5 and 1ng of DNA input.

INSTRUMENT(S): Ion Torrent PGM

ORGANISM(S): Homo sapiens

SUBMITTER: L. Miguel Martins

PROVIDER: E-MTAB-5718 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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