DMT-dT-CE Phosphoramidite丨CAS 98796-51-1

DMT-dT-CE Phosphoramidite丨98796-51-1 is a chemical compound used in DNA synthesis. It is a phosphoramidite derivative of 5'-O-DMT-2'-deoxythymidine, where DMT (dimethoxytrityl) is a protective group, and CE (cyanoethyl) is used as a protecting group for the phosphate backbone. The compound plays a crucial role in the synthesis of DNA oligonucleotides in laboratory settings, particularly in automated solid-phase synthesis.

DNA Oligonucleotide Synthesis

Automated DNA Synthesis: DMT-dT-CE Phosphoramidite丨98796-51-1 is widely used in the automated solid-phase synthesis of DNA oligonucleotides. The presence of the DMT and CE protecting groups makes this compound compatible with the automated coupling and deprotection cycles used in synthesizing custom DNA sequences. The deprotection steps during synthesis allow for the stepwise elongation of DNA chains.

Custom Oligonucleotides: This compound is essential in the production of synthetic oligonucleotides used in various research applications, such as PCR primers, probes, and gene synthesis. Custom DNA sequences are synthesized by adding different phosphoramidites (such as DMT-dT-CE) for each nucleotide in the sequence.

Gene Synthesis

Gene Assembly: DMT-dT-CE phosphoramidite is used in gene synthesis techniques, where short DNA sequences (oligonucleotides) are synthesized and then assembled into larger genes. These synthesized genes can be used in gene therapy, protein expression systems, or in the creation of synthetic genomes.

Synthetic Biology: In synthetic biology, custom DNA sequences are often constructed using a combination of synthetic oligonucleotides like DMT-dT-CE phosphoramidite. These sequences can then be inserted into plasmids or other vectors for use in biological systems or for further genetic manipulation.

Next-Generation Sequencing (NGS) Applications

Library Preparation: DMT-dT-CE Phosphoramidite丨98796-51-1 is sometimes used in the preparation of DNA libraries for next-generation sequencing. These libraries consist of DNA fragments that are used to analyze genomes or transcriptomes by sequencing.

Primer Design: The oligonucleotides synthesized with DMT-dT-CE phosphoramidite can be employed as primers in PCR or sequencing reactions, enabling amplification and analysis of specific DNA sequences.

Mutation Studies

Site-Directed Mutagenesis: The synthesized oligonucleotides using DMT-dT-CE phosphoramidite can also be used for site-directed mutagenesis. This technique allows for the introduction of specific mutations into genes, which can be used to study the function of specific amino acids or regions of a protein or gene.

RNA Synthesis and Modification

RNA Probes and Therapeutics: Although primarily used for DNA synthesis, oligonucleotides synthesized with DMT-dT-CE phosphoramidite can also be used as templates for the synthesis of RNA molecules. These RNA molecules can serve as RNA probes for hybridization experiments or as RNA therapeutics for gene silencing or modulation.

Gene Expression and Protein Synthesis

In Vitro Transcription: DNA sequences synthesized with DMT-dT-CE phosphoramidite can be used as templates for in vitro transcription reactions, which produce RNA molecules for downstream applications like protein synthesis or functional RNA assays.

Protein Production: DNA sequences synthesized with DMT-dT-CE phosphoramidite can be inserted into expression vectors and used for the production of recombinant proteins in bacterial, yeast, or mammalian cells.

High-Quality DNA Synthesis

The use of phosphoramidite chemistry ensures that the DNA synthesis process is efficient, and that high-quality, high-purity DNA oligonucleotides are produced. The DMT protection group allows for the precise addition of nucleotides and prevents premature reactions, ensuring that the final product is a pure, full-length oligonucleotide.

Improved Yield and Purity

The cyanoethyl protection of the phosphate backbone provides stability during the synthesis process, helping to avoid side reactions that could result in truncated or unwanted sequences. This leads to higher yield and purity of the synthesized oligonucleotides.

Versatility in DNA Synthesis

DMT-dT-CE phosphoramidite is compatible with automated DNA synthesizers, allowing for the rapid and high-throughput production of DNA oligonucleotides. This versatility is crucial for applications in genomics, proteomics, and molecular biology, where large numbers of DNA sequences may be required.

Customization of DNA Sequences

The phosphoramidite allows for the easy incorporation of deoxythymidine (dT) into custom-designed DNA sequences. This enables researchers to synthesize oligonucleotides with specific lengths and sequences, tailored to their experimental needs.

DMT-dT-CE Phosphoramidite丨98796-51-1 is a vital reagent in the field of DNA synthesis, enabling the precise and efficient construction of custom DNA sequences for use in various applications, including gene synthesis, mutation studies, next-generation sequencing, and RNA synthesis. Its high selectivity, ease of use in automated systems, and compatibility with phosphoramidite chemistry make it indispensable in the production of synthetic oligonucleotides for research, diagnostics, and biotechnology. The compound's use of protective groups (DMT and CE) ensures that DNA synthesis occurs in a controlled manner, offering high yield and purity of the desired products.