A Real-Time PCR Assay for the Simultaneous Detection of Functional N284I and L412F Polymorphisms in the Human Toll-Like Receptor 3 Gene Robert A. Brown,

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A Real-Time PCR Assay for the Simultaneous Detection of Functional N284I and L412F Polymorphisms in the Human Toll-Like Receptor 3 Gene Robert A. Brown, Raymund R. Razonable The Journal of Molecular Diagnostics Volume 12, Issue 4, Pages 493-497 (July 2010) DOI: 10.2353/jmoldx.2010.090122 Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Schematic representation of the LightCycler melting curve (on F2 channel) of a sample with a wild-type TLR3 851 AA gene that peaked at 62.47°C (solid dark line) and a homozygous gene variant 851TT that peaked at 56.86°C (dashed line). The flat line represents the control with no amplified TLR3 gene. The Journal of Molecular Diagnostics 2010 12, 493-497DOI: (10.2353/jmoldx.2010.090122) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Representation of TLR3 gene sequencing for the A851T single nucleotide polymorphism. A: The wild-type gene containing AAC, which encodes for the amino acid aspargine. B: The A851T single nucleotide polymorphism with the codon ATC, which encodes for isoleucine. The Journal of Molecular Diagnostics 2010 12, 493-497DOI: (10.2353/jmoldx.2010.090122) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Schematic representation of the LightCycler melting curve (F3 channel) of a wild-type TLR3 1234 CC sample that peaked at 61.25°C (solid dark line), a homozygous variant 1234 TT that peaked at 57.98°C (dashed line), and a heterozygous sample containing one allele with 1234C and another allele with 1234T, thereby creating a melting curve with a double peak at both 57.98°C and 61.25°C (thin line). The flat line represents the control with no amplified TLR3. The Journal of Molecular Diagnostics 2010 12, 493-497DOI: (10.2353/jmoldx.2010.090122) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 Representation of TLR3 gene sequencing for the C1234T single nucleotide polymorphism. A: The wild-type gene containing the codon CTC, which encodes for the amino acid leucine. B: The heterozygous gene with one allele with the CTC codon and the second allele with the TTC codon, which encode for leucine and phenylalanine, respectively. C: The homozygous C1234T single nucleotide polymorphism, which contains the TTC codon encoding for phenylalanine. The Journal of Molecular Diagnostics 2010 12, 493-497DOI: (10.2353/jmoldx.2010.090122) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions