We provide sequencing on the ABI Prism 3100xl genetic analyzer. This is is a 16-capillary instrument using fluorescence based electrophoresis technology for DNA sequencing and fragment analysis. For the cycle sequencing we routinely use the 3'-BigDye dideoxynucleotide triphosphates labeling chemistry.
The robust Sanger sequencing chemistry, proven through decades of results, employs fluorescence-based cycle sequencing requires a DNA template, a sequencing primer, a thermal stable DNA polymerase, nucleotides (dNTPs), fluorescently labeled 3'-BigDye dideoxynucleotides (ddNTPs), and buffer. The components are combined in a reaction that is subjected to cycles of annealing, extension, and denaturation in a thermal cycler. Thermal cycling the sequencing reactions creates and amplifies extension products that are terminated by one of the four dideoxynucleotides. The ratio of deoxynucleotides to dideoxynucleotides is optimized to produce a balanced population of long and short extension products.
To submit samples for sequencing