dNTPs, NTPs, and Modified Nucleotides

Thermo Fisher Scientific is one of the few primary manufacturers of nucleotides in the industry. Our nucleotides—including dNTPs, NTPs, and modified nucleotides—are synthesized from high-quality raw materials in state-of-the-art production facilities and available in a variety of formats, formulations, and volumes for convenience and flexibility. Review our offerings below for your applications like PCR, cDNA synthesis, oligo synthesis and labeling, in vitro transcription, and many more.

• High purity (>99%) according to HPLC (Figure 1)
• Free from DNase, RNase, and inhibitors of qPCR, PCR, and RT
• Stable for three years at –20°C
• Available in custom formulations and for commercial supply

For deoxyribonucleotide triphosphate (dNTPs), choose from our catalog of individual products (dATP, dCTP, dGTP, dTTP, and dUTP). dNTPs are available in solutions of each individual species or as a mix of all four species. Custom formulations are also available for specific requirements.

All ribonucleotide triphosphate (NTPs) formulations are designed for convenience and flexibility. Individual nucleotides (ATP, CTP, GTP, and UTP) are supplied in concentrations up to 200 mM in Tris or sodium salt. Custom formulations and nucleotide mixes are also available.

Table 1. Specification for dNTPs and NTPs

	dNTPs	NTPs
Purity*	>99%	>99%
Presence of contaminating nucleotides, nucleosides and PCR inhibitors*	None detected	None detected
DNase, RNase, endo- and exodeoxyribonuclease**	None detected	None detected
Presence of human and E.coli DNA***	Negative	Negative
Stability†	3 years at –20°C >100 freeze-thaw cycles	3 years at –20°C >100 freeze-thaw cycles
Applications	qPCR, RT-qPCR PCR, RT-PCR, cDNA synthesis High-fidelity and long-range PCR Isothermal amplification DNA labeling Cloning Sanger sequencing and next-generation sequencing (NGS)	In vitro transcription mRNA synthesis siRNA synthesis RNA amplification Ligation Phosphorylation
	Order now	Order now

* Measured by HPLC (high performance liquid chromatography)
** Confirmed by incubation of labeled DNA or RNA target with NTPs or dNTPs
*** qPCR test for Alu repeats or 23S rRNA gene
† Stability studies assessed pH, HPLC profiles, and PCR performance.

Highly sensitive diagnostic assays and therapeutic regulatory standards require the highest nucleotide quality and batch-to-batch consistency. Our ISO 13485 and ISO 9001 certified dNTPs have a three year shelf life and are produced under stringent, multi-step QC and manufacturing standards, minimizing inconsistencies and errors in the final product. Manufactured with dedicated equipment in a zero-waste site, each production lot is assayed by the most stringent criteria regarding purity and functionality. For scalable, bulk, and flexible packaging, connect with our expert team for custom solutions.

Purity testing assesses production lots for the desired species via HPLC (>99%) and monitors for the presence of nucleosidic byproducts, residual chemicals, or macromolecule contaminants that occur during production and purification processes. We use a variety of methods to assay for overall purity including HPLC, sensitive colorimetric assays, or performance in PCR (Table 5).

Table 5. Purity assessment of nucleotide solutions.

We manufacture nucleotides with greater than 99% triphosphate purity as assessed by HPLC chromatography (Figure 1).

Trace amount of nucleosidic contaminants can significantly affect PCR performance. These include:

• Mono- and diphosphate forms, which can decrease nucleotide incorporation
• Di-deoxy base forms, which can terminate the amplification reaction
• Modified nucleotides, which can decrease the sensitivity of an assay

Residual chemicals used during nucleotide production or purification can interfere with PCR and are often termed “PCR inhibitors”. Pyrophosphate, a residual contaminant that can occur during production, can inhibit real-time PCR at concentrations of 0.1–0.3 mM, by interfering with fluorescence detection [1]. The presence of inorganic pyrophosphate is easily monitored using the Malachite Green Phosphate Assay which is a colorimetric method that detects formation of a complex between Malachite Green, molybdate, and free orthophosphate.

The presence of macromolecules can cause false test results. These include:

• DNase and RNase contaminants can affect cDNA synthesis resulting in false-negative results (Figure 2)
• Nickase and protease activity can compromise the amplification template
• Contaminating DNA from human and bacterial origins can cause false-positive results

To complement analytical tests for quality control, each lot of Thermo Scientific nucleotides is also analyzed using a variety of functional tests including RT-qPCR and in vitro transcription.

Samples of each lot are tested by RT-qPCR (Figure 3). The assay is performed with low template concentration, which allows detection of even minute variations in dNTP performance.

Samples of each lot are tested by in vitro transcription (Figure 4).

Do you need Invitrogen dNTPs or NTPs for an existing protocol? Please see our Invitrogen Nucleotides Collection to find the products you need.

1. Ginzinger, D.G. 2002 Gene quantification using real-time quantitative PCR: an emerging technology hits the mainstream. Experimental Hematology 30:503–512.