ddNTPs (Lithium salt) from GeneON  in combination with modified Taq DNA Polymerase are used as 3'-end chain terminators in Sanger sequencing. @ pH 7,5  in water the high purity between > 98 % (HPLC) offers a great performance in chain termination sequencing.

The nucleotides are especially manufactured and tested for application in sequencing reactions.

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. 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 but 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, but every DNA strand will end at the same type of base (A, C, G or T).

Used in Methods:

- Sanger: dideoxy chain termination sequencing

- single nucleotide polymorphisms (SNPs) genotyping by single base pair extension (SBE)

Molecular Weight and Formula:

ddATP: 475.18 g/mol C10H16N5O11P3 (acid free)

ddCTP: 451.16 g/mol C9H16N3O12P3 (acid free)

ddGTP: 491.18 g/mol C10H16N5O12P3 (acid free)

ddTTP: 466.17 g/mol  C10H17N2O13P3 (acid free)

Concentration: 10 - 11 mM

Spectro-Data @ pH 7,5 ±0.5, Tris-HCL

ddATP: λmax 259 nm, ε 15.1 L mmol-1 cm-1 

ddCTP: λmax 271 nm, ε 8.9 L mmol-1 cm-1

ddGTP: λmax 252 nm, ε 14.2 L mmol-1 cm-1 

ddTTP: λmax 267 nm, ε 9.6 L mmol-1 cm-1

Note: spin all reagents in the vial before opening and after setting up the Termination Mix

Transport: on blue ice or ambient temperature

Storage: few days at room temperature; for long term storage @ -20 °C

Quality tests:
The ddNTPs are tested:
- for the absence the absence of DNases and RNases
- tested in chain termination sequencing reaction
- produced in German factory (ISO: 9001/2001)