The use of a sulfurizing reagent during the regular synthesis cycle using phosphoramidite chemistry has revolutionized the production of phosphorothioate oligonucleotide analogues. Undoubtedly, this ease of preparation of phosphorothioates has made this oligonucleotide modification by far the most common in research. Glen Research was one of the first sources of the sulfurizing reagent, 3H-1,2-benzodithiol-3-one 1,1-dioxide, popularly known as Beaucage Reagent (1).1 This sulfurizing reagent has found common use in the face of a plethora of rival reagents over the years because of its high efficiency, fast reaction time, and widespread availability. The one mild flaw we have found with Beaucage Reagent is that, although it is quite stable in acetonitrile solution in a silanized amber bottle, it is has relatively poor stability in solution once installed on the DNA synthesizer. Consequently, we have not been able to supply a sulfurizing solution, preferring to supply the powdered reagent along with an appropriate silanized bottle. The customer then weighs an appropriate amount of reagent into the silanized bottle and adds acetonitrile at a concentration of 1g/100mL. Over the years, we have considered other sulfurizing reagents but we were not able to find another reagent that exhibits the same fast sulfurization kinetics along with improved stability on the synthesizer. RNA Sulfurization
DNA oligomers are the foundation of the process. The essential feature of DNA synthesis is that no naturally isolated DNA is used. This step deviates from the traditional method of studying genetics, by which a preexisting genetic sequence was extracted from an organism to be amplified and later studied. With gene synthesis we, at Synbio Technologies, are capable of generating de novo genetic sequences that may not currently be present anywhere within nature. Although clonal plasmid-based intermediates might exist during the assembly of a target DNA, every base originates as a phosphoramidite molecule at the beginning of DNA synthesis. Today, all DNA synthesis methods begin with solid-phase phosphoramidite chemistry to construct single-stranded DNA molecules ranging in size between 10 and 100 base pairs in length. These single stranded molecules are then enzymatically assembled into larger molecules. This process is then repeated until the required sequence length is generated. This process has been perfected by Synbio Technologies through our three phase Syno Platform. This platform offers the ability to generate sequences up to and including 200kb in length with one hundred percent accuracy, guaranteed to be identical to the sequence requested by the customer.
"This is how the synthesis of new molecules …
The most common usage for oligonucleotide phosphorothioates has been in the production of antisense oligodeoxynucleotides destined for use in identifying or modifying gene expression. Now, phosphorothioate linkages are popping up in the RNA world and sulfurizing RNA linkages with reagents like Beaucage Reagent has proved to be much more difficult than DNA linkages. The phosphorothioate (PS) linkage is a not-so-expensive way of increasing the stability of nucleic acids and increasing nuclease resistance of RNA. Now, it has been shown2 that fully PS oligos can promote the delivery of siRNA in cell culture. This siRNA uptake is sequence-independent and the length seems to vary between 30 and 70 nucleotides depending on the cell line. Even though this method is not yet as efficient as the cationic lipids, it opens the way to possible new methods. Reasons that may explain this are not understood at this time.
Small Molecule DNA and RNA Binders: From Synthesis to Nucleic Acid ..
Synbio Technologies is a DNA technology company whose main focus is . This is seen in our high quality products generated by Synbio Technologies’ proprietary Syno® DNA synthesis platform which can synthesize sequences up to and including 200kb in length. This length is characteristic of DNA sequences which make up genes, genomes, and various biological pathways. In addition to this, the professional Syno®Codon software can significantly optimize the original sequences from customers, and provide synthesized DNA sequences with one hundred percent accuracy within 5 days. It is these factors that have led Synbio Technologies to become one of the premier companies within the gene synthesis industry.
the synthesis of new double helix molecules.
Synbio Technologies is building up the first integrated GPS (Genotype, Phenotype and Synotype) system aimed to a quick and easy translation or reverse translation between "Genotype" and "Phenotype" by using our proprietary "Synotype" platform. The company's scientific capabilities encompass areas such as DNA engineering, DNA synthesis, genome synthesis, pathway synthesis, synthetic biology, pharmacogenomics, microbiology, translational biology and the applications of synthetic biology. Synbio Technologies' team has a proven track record regarding translating scientific breakthroughs into cost effective biological solution.
Processes of DNA Replication and Protein Synthesis ..
DNA polymerases cannot initiate synthesis of a parental chain using a free dNTP(N=A, G, C, and T) as the first substrate. They have to add dNTPs onto an RNA primer or an existing DNA segment. The primer provides a free 3'-OH group for the incoming nucleotide to form a phosphodiester bond. The dNTP is added as its monophosphate residue, leaving pyrophosphate as the other product of the reaction (Fig. 2, below).