C677T Single Nucleotide Polymorphisms of the Human Methylene Tetrahydrofolate Reductase and Specific Identification

Abstract

We present an alternative method to those commonly used in genotyping. Two-color cross-correlation FCS allows the detection of the fluorescence signals specifically associated with the heterozygous mutated, the homozygous mutated, and normal individuals, as exemplified in this study. The presence of nonspecific amplification products, which interfere with subsequent DNA analysis, could therefore highlight the need for two-color cross-correlation FCS as a means of discriminating between specific association of the fluorescence signals with the target DNA and DNA not related to the target.

Direct Quantification of Gene Expression Using Fluorescence Correlation Spectroscopy

“…Therefore, an excess of free probes does not contribute to cross-correlation. This advantage of fluorescence cross-correlation spectroscopy (FCCS) made it possible to quantify the methylene tetrahydrofolate reductase-genotypes at site C677T (the heterozygous mutated, the homozygous mutated, and the normal) directly on unamplified genomic DNA [14,19]. Fig.…”

“…In the first case, all of these methods employ slightly different approaches, but the underlying principle is the same in that polymerase chain reaction [14][15][16][17] or other amplification method is used [18]. In one of the first applications of FCS for detecting genomic DNA, lambda phage DNA amplified with asymmetric PCR was detected [17].…”