Deoxyribonucleoside Triphosphates
Deoxyribonucleoside Triphosphates - Related Videos
15:22
Nucleoside Triphosphates - From Synthesis to Biochemical Characterization
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Cited by 4 •
2014
The protocol described herein aims to explain and abridge the numerous obstacles in the way of the intricate route leading to modified nucleoside triphosphates. Consequently, this protocol facilitates both the synthesis of these activated building-blocks and their availability for practical applications.
02:40
DNA Replication
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2020
DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied. After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells. Replication in Prokaryotes DNA replication uses a large number of...
13:19
Chemical Triphosphorylation of Oligonucleotides
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Cited by 1 •
2022
Oligonucleotide 5′-triphosphates are ubiquitous components in essential biological pathways and have seen increasing use in biotechnology applications. Here, we describe techniques for the routine synthesis and purification of oligonucleotide 5′-triphosphates, starting from oligonucleotides prepared by standard automated synthesis techniques.
06:16
Preparation Of Gushukang (GSK) Granules for In Vivo and In Vitro Experiments
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Cited by 4 •
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This article provides a detailed protocol for preparing a working solution of Gushukang granules for animal studies and GSK granule containing serum for in vitro experiments. This protocol can be applied to pharmacological investigations of herbal medicines as well as prescriptions for both in vivo and in vitro experiments.
12:12
Analysis of Termination of Transcription Using BrUTP-strand-specific Transcription Run-on (TRO) Approach
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Cited by 7 •
2017
We describe a basic experimental approach for analysis of termination of transcription by RNA polymerase II in vivo using BrUTP by the strand-specific transcription run-on (TRO) approach in budding yeast. This protocol can be extended to study transcription termination by other RNA polymerases both in yeast and higher eukaryotes.
FAQs
DNA polymerase selects among dATP, dCTP, dGTP, and dTTP according to complementarity with the template sequence. This selection determines which base becomes part of the growing DNA strand, so the chemical recognition step directly converts template information into a corresponding nucleotide sequence. The process therefore links molecular structure with the accuracy of genetic information transfer.
During incorporation, DNA polymerase transfers the dNTP’s 5′ phosphate to the growing strand’s 3′ hydroxyl, forming a phosphodiester bond and releasing pyrophosphate. This reaction couples strand extension with cleavage of the incoming substrate’s triphosphate group. Consequently, dNTPs provide not only the nucleotide added to DNA but also the chemical energy associated with bond formation.
The four substrates differ in their nitrogenous bases: dATP, dCTP, dGTP, and dTTP. Their identities determine which complementary base is incorporated at each position, while their shared deoxyribose and triphosphate features support the same general polymerization chemistry. Varying the order of these substrates produces the sequence diversity required for DNA to encode genetic information.
Controlled use of dNTPs connects the immediate chemistry of DNA synthesis with broader biological regulation. Polymerase activity depends on selecting the appropriate substrate from the available nucleotide pool, while studies of nucleotide metabolism examine how these building blocks are managed. Their controlled use also supports investigations of replication fidelity, linking substrate handling to accurate genome copying.
In PCR, dNTPs serve as the substrates that DNA polymerase incorporates while producing amplified DNA. Each of the four substrates is needed to represent the possible bases in the product sequence, and their triphosphate groups support phosphodiester-bond formation. Their use makes PCR a practical application of the same polymerization chemistry that underlies DNA replication.
DNA sequencing and cloning both rely on controlled DNA synthesis, making dNTPs central reagents in these applications. During synthesis, polymerase uses the available dATP, dCTP, dGTP, and dTTP to generate DNA with a defined base sequence. The resulting products can provide sequence information in sequencing or contribute to the construction of recombinant DNA in cloning.
Molecular diagnostics use DNA synthesis to detect or analyze genetic information, so dNTPs supply the substrates required for polymerase-based reactions. From a chemistry perspective, they illustrate how sugar structure, base identity, and a reactive triphosphate group combine in one molecule. Studying these features supports research on DNA synthesis, replication fidelity, and nucleotide metabolism.