Allelic discrimination by nick-translation PCR with fluorgenic probes

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ID: 305634
1993
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Abstract
Nick-translation PCR was performed with fluorogenic probes. Two probes were used: one complementary to a sequence containing the F508 codon of the normal human cystic fibrosis (CF) gene ( wt DNA) and one complementary to a sequence containing the AF508 three base pair deletion ( mut DNA). Each probe contained a unique and spectrally resolvable fluorescent Indicator dye at the 5′ end and a common quencher dye attached to the seventh nucleotide from the 5′ end. The F508/ΔAF508 site was located between the indicator and quencher. The probes were added at the start of a PCR containing mut DNA, wt DNA or heterozygous DNA and were degraded during thermal cycling. Although both probes were degraded, each probe generated fluorescence from its indicator dye only when the sequence between the indicator and quencher dyes was perfectly complementary to target. The identity of the target DNA could be determined from the post-PCR fluorescence emission spectrum.
Reference Key
openalex_W1988966853 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Linda G. Lee, Charles R. Connell, Will Bloch
Journal Nucleic Acids Research
Year 1993
DOI
10.1093/nar/21.16.3761
URL
https://doi.org/10.1093/nar/21.16.3761
Keywords Keywords not found

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