AAT Bioquest


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Physical properties
Molecular weight576.24
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure


Molecular weight
5-Propargylamino-3'-azidomethyl-dUTP is a key building block for preparing fluorescent conjugates that are used in the next generation sequencing (NGS). NGS uses a similar chain termination method to the earlier Sanger sequencing, but NGS is carried out by fluorescence-labeled nucleotide analogs acting as reversible terminators of the amplification reaction. NGS relies on the blockade of DNA polymerization that is reversible while the Sanger sequencing uses the irreversible blockade of DNA polymerization by ddNTPs. Another different feature of NGS is that the clonal amplification in vitro to multiply the number of molecules to be sequenced is conducted by means of bridge PCR. In this platform, the fragments are joined to primers immobilized on a solid surface, performing an amplification in situ, generating clusters of DNA with identical molecules. In each cycle, the four nucleotides of reversible termination are simultaneously added and incorporated by the polymerase they complement. These nucleotides are chemically blocked—by substituting the 3′-OH group for a 3′-o-azidomethyl group—to prevent the polymerase from incorporating more than one nucleotide in each cycle. Upon incorporation of a nucleotide, a fluorescence signal is measured in different channels for different bases. Concerning the next cycle, the nucleotides that have not been incorporated are washed and the chemical blockade of the 3′ end is removed with TCEP. Once the fluorescence signal is collected, a new cycle begins, repeating this dynamic until the sequencing of each fragment is finished. In summary, the NGS sequencing reaction is carried out in three steps: addition of nucleotides, imaging, and regeneration of 3′-OH by fluorophore cleavage.


Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of 5-Propargylamino-3'-azidomethyl-dUTP to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM173.539 µL867.694 µL1.735 mL8.677 mL17.354 mL
5 mM34.708 µL173.539 µL347.078 µL1.735 mL3.471 mL
10 mM17.354 µL86.769 µL173.539 µL867.694 µL1.735 mL

Molarity calculator

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View all 50 references: Citation Explorer
Current scenario of the genetic testing for rare neurological disorders exploiting next generation sequencing.
Authors: Di Resta, Chiara and Pipitone, Giovanni Battista and Carrera, Paola and Ferrari, Maurizio
Journal: Neural regeneration research (2021): 475-481
Application of Next Generation Sequencing in Laboratory Medicine.
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Journal: Annals of laboratory medicine (2021): 25-43
Amplicon-Based Next-Generation Sequencing for Detection of Fungi in Formalin-Fixed, Paraffin-Embedded Tissues: Correlation with Histopathology and Clinical Applications.
Authors: Larkin, Paige M K and Lawson, Katy L and Contreras, Deisy A and Le, Catherine Q and Trejo, Marisol and Realegeno, Susan and Hilt, Evann E and Chandrasekaran, Sukantha and Garner, Omai B and Fishbein, Gregory A and Yang, Shangxin
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NGS_SNPAnalyzer: a desktop software supporting genome projects by identifying and visualizing sequence variations from next-generation sequencing data.
Authors: Lee, Dong-Jun and Kwon, Taesoo and Kim, Chang-Kug and Seol, Young-Joo and Park, Dong-Suk and Lee, Tae-Ho and Ahn, Byung-Ohg
Journal: Genes & genomics (2020)
Identification of the novel HLA-C*06:297 allele by next generation sequencing.
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The Impact of Next-Generation Sequencing on the Diagnosis, Treatment, and Prevention of Hereditary Neuromuscular Disorders.
Authors: Beecroft, Sarah J and Lamont, Phillipa J and Edwards, Samantha and Goullée, Hayley and Davis, Mark R and Laing, Nigel G and Ravenscroft, Gianina
Journal: Molecular diagnosis & therapy (2020)
Identification of the novel HLA-DQB1, allele DQB1*06:02:44 by next-generation sequencing.
Authors: Ananeva, Anastasiia and Osipova, Nailia and Andryushkina, Anna V and Shagimardanova, Elena I
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Detection of Listeria monocytogenes in a patient with meningoencephalitis using next-generation sequencing: a case report.
Authors: Lan, Zi-Wei and Xiao, Min-Jia and Guan, Yuan-Lin and Zhan, Ya-Jing and Tang, Xiang-Qi
Journal: BMC infectious diseases (2020): 721
Author Correction: Ultrasensitive amplicon barcoding for next-generation sequencing facilitating sequence error and amplification-bias correction.
Authors: Ahmed, Ibrahim and Tucci, Felicia A and Aflalo, Aure and Smith, Kenneth G C and Bashford-Rogers, Rachael J M
Journal: Scientific reports (2020): 17010
PCAT18, as a novel differentially regulated long noncoding RNA in adult acute myeloid leukemia patients revealed by next-generation sequencing.
Authors: Zhang, Jihong and Zhang, Henan and Wang, Xiaohui and Zhao, Yue and Fu, Yu and Liu, Xuan
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