Single Reaction Sanger Sequencing

Sanger Sequencing for Single Targets

Labeled dideoxynucleotide termination of DNA strand growth, pioneered by Nobel Laureate Frederick Sanger, has become the cornerstone of Genomics. The ABI 3730 utilizes lasers to illuminate the fluorescent dyes on the terminated strand. Single Reaction Sequencing has grown to Massively Parallel Sequencing, a.k.a., Next Generation Sequencing. Illumina’s platforms use the same technology, but the fluorescence is read directly through a sensitive light meter. Thermofisher’s NGS technology now has a silicon chip with millions of pH meters built in, but doesn’t measure chain termination, but a H+ release from a growing strand, very different from Sanger Sequencing. Although powerful, NGS covers full genomes or large DNA populations, many researchers still need to focus on a small set of genes. Sanger Sequencing on single targets is still a very viable technology in the lab, including clinical work.

Utilizing a single target and a single primer, floursceinated dideoxynucleotides terminate a growing strand of DNA in a thermal cycling reaction. After washing, the DNA population from the cycle sequencing reaction are separated by size in capillary electrophoresis then the dideoxynucleotides are illuminated with a laser.

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  • Up to 1100 bp per Reaction
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  • 17 Universal Primers Available
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