Used in single nucleotide polymorphisms and as 3´-end chain terminators in Sanger sequencing.Stringent QC at ISO-certificated BULK-Production.
ddNTPs in combination with modified Taq DNA Polymerase are used as 3′-end chain terminators in Sanger sequencing. @ pH 8,0 in water the high purity between 98 – 99 % (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 Sange. 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).
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 day at room temperature; for long term storage @ -20 °C
Quality tests:
The ddNTPs are tested:
– for the absence the absence of DNases and RNases
– for succesfully sequencing reaction
– produced in German factory (ISO: 9001/2001)
Standard protocol for 50 µl “Termination” Mix:
| Component | Volume | final concentration |
| dNTP’s (1 mM each) | 5 µl of each dNTP | 400 µM |
| ddNTP’s (10 µM) | 2 µl | 0,4 mM |
| Water | 25 µl | |
| Total volume | 50 µl |
| Product | Description |
|
1910-020 |
Set of ddNTP 4×10 mM 4 x 200 μl (4 x 2 μmol) |
| 1910-025 |
Set of ddNTP 4×10 mM 4 x 1000 μl (4 x 10 μmol) |
| 1910-020A |
ddATP 10 mM 200μl (2 μmol) |
| 1910-025A |
ddATP 10 mM 1000μl (10 μmol) |
| 1910-020C |
ddCTP 10 mM 200μl (2 μmol) |
| 1910-025C |
ddCTP 10 mM 1000μl (10 μmol) |
| 1910-020G |
ddGTP 10 mM 200μl (2 μmol) |
| 1910-025G |
ddGTP 10 mM 1000μl (10 μmol) |
| 1910-020T |
ddTTP 10 mM 200μl (2 μmol) |
| 1910-025T |
ddTTP 10 mM 1000μl (10 μmol) |
Datasheet
Stability Test Report
Material Safety Datasheet
