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Next Generation Sequencing

CYTOP® 208 – one of the preferred reducing agents showing exceptional stability at biological pH, for the improved performance rapid reduction of a wide range of differentially functionalized small-molecule disulfides in the SBS process.

Next generation sequencing (NGS) is used for DNA and RNA sequencing and variant or mutation detection.(1) NGS enables rapid sequencing of thousands of genes leading to its use for disease diagnosis and clinical decision making. It can also play a role in the realization of personalized precision medicine.

Next Generation Sequencing and SARS-CoV-2

During the pandemic, NGS has been utilized to track the origins of outbreaks, both identify, and map the whole genome, of new variants, facilitate the development of tests, and contribute to the development of vaccines and treatments.(2) GISAID (gisaid.org), the Global Initiative on Sharing All Influenza Data has received >7 million SARS-CoV-2 genome sequences in its database.

Sequencing by Synthesis

The most widely used NGS technology is sequencing by synthesis (SBS). SBS involves the generation of millions of DNA reads in a single sequence run, the typical steps are distribution of single DNA molecules, amplification, DNA synthesis reactions with labeled nucleotides and imaging / detection.(3) One of the detection methods is the use of modified nucleotides with different fluorescent labels, corresponding to each of the four bases, during the synthesis reactions. The modified nucleotides are removed before the next round of synthesis.(4)

Application of CYTOP® 208 (Strem#: 97-1002) in Sequencing by Synthesis

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The modified nucleotides used in SBS are often attached by the formation of disulfide bonds. Once the tag has been read these labeled nucleotides are removed by cleaving the sulfur-sulfur bonds. CYTOP®208  (Strem#: 97-1002) is one of the preferred reducing agents for this transformation. The reagent shows exceptional stability at biological pH under which condition it permits the rapid reduction of a wide range of differentially functionalized small-molecule disulfides with improved performance in comparison with the alternative reagents TCEP and DTT.(5)

CYTOP® 208 (Strem#: 97-1002) is a commercial scale, relatively air-stable, water soluble tertiary phosphine that is stable under both acidic and basic conditions.

To learn more about other specialty phosphorus products, please refer to the Solvay offering at Strem or the Solvay website.

References:

  1.       Cancer Biol. Med. 2019, 16, 4-10.
  2.       Briefings in Bioinformatics. 2021, 22, 616-630.
  3.       Curr. Protoc. Mol. Biol. 2018, 122.
  4.       World patent application 2021/067970.
  5.       Bioorganic Medicinal Chemistry Letters, 2015, 25, 4114-4119.

 

Featured Product:

97-1002 Tris(3-hydroxypropyl)phosphine, min. 84% [CYTOP® 208 ORGANOPHOSPHINE](4706-17-6)

 

Related Products:

All products sold in collaboration with Solvay
Phosphorus Ligands and Compounds Booklet

 

 

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