DDNTP SET SEQUENCING GRADE

General description

The Dideoxynucleoside Triphosphate Set, Sequencing Grade, sodium salt solution, consists of dideoxynucleotides of very high purity (ddNTP HPLC, area% ≥98%; ddNDP HPLC, area% ≤1.5%), specifically manufactured and tested for sequencing reactions.
The set contains 4 individual vials of ddATP, ddCTP, ddGTP, and ddTTP, each at 10 mM as a clear, colorless solution of the sodium salt (pH 8.3).

Application

2′,3′-Dideoxynucleoside triphosphates inhibit the chain elongation of a given primer catalyzed by the DNA polymerase (e.g., Klenow enzyme) and are therefore used for DNA sequencing according to Sanger (Sanger, F. et al. (1977) PNAS USA 74, 5463).
Sequencing is achieved by including in each reaction a dideoxynucleotide that acts as a chain terminator. Four reactions are set up, each containing the same template and primer, and a chain terminator specific for A, C, G, or T. Because only a small amount of the chain terminator is included, incorporation into the new DNA strand is a random event. Each reaction therefore generates a collection of fragments, and every DNA strand will end at the same type of base (A, C, G, or T).

Packaging

The set contains 4 individual vials of ddATP, ddCTP, ddGTP and ddTTP each at 10 mM as a clear, colorless solution of the sodium salt (pH 8.3).

Quality

Each lot of dideoxynucleotide solution is assayed for function in sequencing reaction.
Tested for absence of contaminating deoxyribonucleases, ribonucleases and nicking activities according to the current Quality Control procedures.

Preparation Note

Activator: Mg²⁺, Co²⁺ and Mn²⁺ ions
Working solution: Preparation of Termination Mix
Before preparing the termination mix, dilute an appropriate volume of each dideoxynucleotide in water to a concentration of 10 μM.

Other Notes

For life science research only. Not for use in diagnostic procedures.