Methods

GTCA Testing Methods

Not all samples are amenable to genetic testing using the same methods.  To ensure you receive the most accurate results we employ a number of methods as appropriate for your samples.

Melt curve analysis

The longer the amplicon the more hydrogen bonds present

Assays are designed to amplify specific regions of the target sequence in the presence of SYBR green. Post amplification, the amplicons are subjected to an increasing temperature while data is collected. SYBR green intercalates with double-stranded DNA and releases light at a wavelength detected by the instrument. As the temperature increases, the double-stranded DNA will melt apart based primarily on two factors.

  1. The length of the DNA amplicon. The longer the amplicon the more hydrogen bonds present and the greater the required energy to separate the strands.
  2. The G-C content.

This is due to the fact that G-C pairs share 3 hydrogen bonds, while A-T pairings only share 2 hydrogen bonds.  Therefore, amplicons designed for this method can be differentiated based on the melt profiles of these different products.

This is due to the fact that G-C pairs share 3 hydrogen bonds, while A-T pairings only share 2 hydrogen bonds.  Therefore, amplicons designed for this method can be differentiated based on the melt profiles of these different products.

Melt curve analysis

The longer the amplicon the more hydrogen bonds present

Assays are designed to amplify specific regions of the target sequence in the presence of SYBR green. Post amplification, the amplicons are subjected to an increasing temperature while data is collected. SYBR green intercalates with double-stranded DNA and releases light at a wavelength detected by the instrument. As the temperature increases, the double-stranded DNA will melt apart based primarily on two factors.

  1. The length of the DNA amplicon. The longer the amplicon the more hydrogen bonds present and the greater the required energy to separate the strands.
  2. The G-C content.

This is due to the fact that G-C pairs share 3 hydrogen bonds, while A-T pairings only share 2 hydrogen bonds.  Therefore, amplicons designed for this method can be differentiated based on the melt profiles of these different products.

This is due to the fact that G-C pairs share 3 hydrogen bonds, while A-T pairings only share 2 hydrogen bonds.  Therefore, amplicons designed for this method can be differentiated based on the melt profiles of these different products.