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ddNTPDideoxynucleotide Triphosphate
ddNTPDideoxyribonucleotide triphosphate
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References in periodicals archive ?
(36) Dideoxynucleotide sequencing is based on the synthesis of new DNA strands (complementary to the target sequence of interest) in the presence of primers, deoxynucleotide triphosphates (dNTPs) and dideoxynucleotide triphosphate chain terminators (ddNTPs).
After the reaction, 5'-phosphoryl groups of unincorporated dideoxynucleotide triphosphates were removed by addition of 1.0 U of calf intestinal phosphatase (Finnzymes) and incubation of the samples at 37[degrees]C for 1 h and at 75[degrees]C for 15 min to successively deactivate the enzyme.
Four reaction tubes were each prepared with single-stranded DNA template (2.5 [micro]l) for the sequence of interest, radiolabeled primers (1 [micro]l), dideoxynucleotide triphosphates and DNA polymerase (9.5 [micro]l), and four deoxynucleotide triphosphates.
Quantification of MALDI-TOF-MS signals requires low background and clear peak separation; therefore, use of deoxynucleotide triphosphates and dideoxynucleotide triphosphates for addition of one or two bases, respectively, producing mass differences of ~300 Da, is a prerequisite for reproducible results.
This mutation permits efficient incorporation of dideoxynucleotide triphosphates (ddNTPs), which usually are highly discriminated against by standard wild-type Taq DNA polymerase (5).
The 11 purified primers (Genosys; Table 1), each specific for a screened mutation, bind to the complementary sequence in the presence of fluorescently labeled dideoxynucleotide triphosphates (ddNTPs), and the AmpliTaq DNA Polymerase extends the primer by adding a single ddNTP to its 3' end.