RESUMO
It has been very difficult to detect and quantify multiple somatic mutations simultaneously in single-tube qPCR. Here, a novel method for simultaneous detection of multiple mutations and a melting curve analysis was developed by using clamping PNA and detection PNA probes. Each PNA probe was designed to have a specific melting temperature by the introduction of γ-PNA monomer. This technique was successfully applied to the detection of six genotypes in two different mutations of K-RAS at the same time. Such simultaneous analysis of an amplified curve and a melting curve in qPCR can be widely used for the early diagnosis of cancer and determining the prognosis of drug treatments.
Assuntos
Genes ras/genética , Mutação , Ácidos Nucleicos Peptídicos/genética , Reação em Cadeia da Polimerase/métodos , Alanina/química , Linhagem Celular , Dimerização , Técnicas de Genotipagem/métodos , Células HeLa , Humanos , Lisina/química , Sondas Moleculares , TemperaturaRESUMO
Synthesis of self-activated peptide nucleic acid (PNA) monomers and an efficient method for PNA synthesis using a benzothiazole-2-sulfonyl (Bts) group as an amine-protecting group as well as an acid-activating group are reported. Couplings were complete within 120 min, and the deprotection was performed in 10 min. This Bts strategy provides a high purity PNA oligomer and is appropriate for large-scale synthesis. The results of the 15-mer PNA oligomer are described.
Assuntos
Ácidos Nucleicos Peptídicos/síntese química , Amidas/química , Técnicas de Química CombinatóriaRESUMO
We present a way to improve the dispersion tolerance of an electrical-binary-signal-based duobinary transmitter, implemented by using a dual-arm Mach-Zehnder modulator driven with two complementary binary signals. Successful transmission over 200 km of single- mode fiber is achieved by optimizing the relative time delay between the binary signals and the driving voltage.
RESUMO
Peptide nucleic acids (PNAs) are DNA analogues in which the nucleic acid backbone is replaced by a pseudopeptide backbone and nucleobases are attached to the backbone by methylene carbonyl linkers. γ-Carbon modification of the PNA structure allows monomers, and subsequently oligomers, with improved properties to be obtained. In this study, we report the convenient synthesis of γ-lysine-modified PNA monomers for pyrimidine bases (thymine and cytosine) with high optical purity (> 99.5%) and direct enantiomer separation of γ-lysine-modified PNA analogs, using chiral HPLC to determine the optical purity.