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

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

PROVIDER: PRJEB20590 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR1950780.fastq.gz	Fastqsanger.gz
	ERR1950790.fastq.gz	Fastqsanger.gz
	ERR1950800.fastq.gz	Fastqsanger.gz
	ERR1950781.fastq.gz	Fastqsanger.gz
	ERR1950791.fastq.gz	Fastqsanger.gz

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

Project description: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.

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