Set of ddNTP (ddATP, ddCTP, ddGTP, ddTTP; 5µM/each)
Opis
Ultrapure ddNTPs as set of. 4 x 1ml
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)
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).
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
Stability tests / Quality Tests
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)
Dane techniczne
Opakowanie | zestaw |