Nonstandard abbreviations: ddPCR, droplet digital PCR; FDA, Food and Drug Administration; EC, European Conformity; NSCLC, non-small cell lung cancers; ctDNA, circulating tumor DNA; WT, wild-type; FFPE, formalin-fixed and paraffin embedded; CRC, colorectal cancer; cfDNA, cell-free DNA; gDNA, genomic DNA; NGS, next-generation sequencing; PBMC, peripheral blood mononuclear cells; MAF, mutant allele frequency; REF, reference; NFQ
, nonfluorescent quencher; LOB, limit of blank; LOD, limit of detection.
Cook et al., 2010 PSQI; CAPS; NFQ
;NES; PSQI-A; PCL-M; BDI; ns 36-ISFHS.
The sequences were as follows: forward primer 5'-GCA CCT GAC TCC TGA GGA GAA-3', reverse primer 5'-CAC CAA CTT CAT CCA CGT TCA-3', and a single-labeled fluorescent MGB-probe 5'-FAM-TCT GCC GTT ACT GCC CT-MGB-NFQ, where MGB is a minor groove binding molecule and NFQ
a nonfluorescent quencher molecule.
, nonfluorescent quencher; T*, A*, G*, super T base, super A base, super G base (Nanogen); Z*, neutral base (Nanogen).
(4) Nonstandard abbreviations: RTD PCR, real-time detection PCR; WB, whole blood; [T.sub.m] melting temperature; FVL, factor 5 Leiden; PT, prothrombin; HFE, hemochromatosis gene; FRET, fluorescence resonance energy transfer; MGB, minor groove binding; NFQ
, nonfluorescent quencher; and 6-FAM, 6-carboxyfluorescein.
The sequence of the reporter probe was 5'-(FAM)TCTGCGTAGGCAATCC(NFQ
)-3' (FAM, 6-carboxyfluorescein; NFQ
, nonfluorescent quencher; complementary to nt 17,745-17,760).
A nonfluorescent quencher (NFQ
) and the MGB were added to the 3' end.
The 5'-fluorescently labeled probes [6-carboxyfluorescein (FAM) or VIC] were conjugated to a nonfluorescent quencher (nfq
) and an MGB group at the 3' end.