Category: DNA synthesis

Introduction of A New DNA Synthesis Method

Gene synthesis is the main content of current synthetic biology. Through gene synthesis, genes that do not exist in nature can be obtained, which provide a whole new direction for humans to transform organisms. In the foreseeable future, gene synthesis will play an important role in the field of life sciences. However, the current speed of gene synthesis still cannot fully meet the needs of biological development. A few days ago, a new document published by “Nature Biotechnology” pointed out that a new gene synthesis technology actually shortened the synthesis time of new genes to only one day!

Traditional gene synthesis is mostly used for the creation of antibody genes, primarily using terminal deoxynucleotide transferase (TdT), which can add new nucleotides to the oligonucleotide chain without following the DNA template chain. Thereby, new variants of millions of antibody genes have been written, and the immune system can use these variants to target invaders. But, natural TdT can only randomly add new deoxynucleotides, and researchers currently cannot accurately control the synthesis process.

gene-synthesis

DNA sequence synthesis process under the action of TdT

The researchers of this article designed the TdT-dNTP conjugate, which can be actively covalently linked to the 3’end when incorporated into the primer. This prevents the binding of other TdT-dNTP conjugates to the primer. During further extension, the linker can be cleaved, the 3’end of the primer is deprotected, and a single nucleotide is added to the extension. This makes the “termination” process reversible from now on, and researchers can turn on or end the role of TdT as required.

The researchers have shown that the active TdT-dNTP conjugate can connect the nucleoside triphosphate to various points on the surface of the polymerase through two different linkers. They found that the TdT-dNTP conjugate can achieve DNA in 10-20s within one single nucleotide extension of the molecule. So, this cycle of extension and deprotection reaction can quickly and accurately write a defined sequence, and it takes only one day to synthesize a complete gene.

dna-synthesis

TdT-dNTP achieves single nucleotide extension of DNA molecules within 10-20s

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References

Palluk S , Arlow D H , Rond T D , et al. De novo DNA synthesis using polymerase-nucleotide conjugates[J]. Nature Biotechnology, 2018, 36(7).

Gene synthesis in synthetic biology

Gene synthesis is a kind of technique that has been used to synthesize double-stranded DNA molecules in vitro by artificial ways. It is different from the single-stranded oligonucleotide synthesis in term of the sequence length. The longest synthesized oligonucleotide fragment is just up to 100nt. However, double-stranded DNA molecules, as products of the gene synthesis, can reach 50bp-12 kbp in length. Besides, compared with the approach of gaining genes from existing organisms, gene synthesis do not need templates, so it is not restricted by gene donors.

The first artificially synthesized gene appeared in the 1960s. As a main subject in current synthetic biology, gene synthesis sets a new direction for human beings to change organisms in gene level. It is predictable that in the near future gene synthesis will play a significant role in life science. Moreover, its advantages in new energy, new materials, artificial organisms, DNA vaccines, and biomedicine, have initially been noted by scientists.

Synbio Technologies provides our customers comprehensive DNA solutions that can satisfy all their DNA manufacturing needs. Our Syno® DNA synthesis platforms provide gene and fragment synthesis, high-throughput DNA synthesis, pathways synthesis, genome assembly, etc. Our proprietary algorithm based on DNA sequence complexity index (CI) system can precisely facilitate the success of complex sequences, for example: repeated sequences, high/low GC content, hairpin and poly structure, ultra-long sequences (up to 150 kb), etc. We have developed the most complete platforms of oligo synthesis, gene synthesis (fragment and clonal genes) and next-generation high-throughput gene synthesis to meet our customers’ needs in quality, speed, budget, and applications.

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High-throughput gene synthesis

Synthetic biology research began with the redesign, reconstruction, and optimization of existing life systems, so that the transformed biological system can meet the purposeful engineering applications of human beings. It has now changed from a single gene research model to integrated modules and loops, based on multi-gene and systemic research on pathways (even genome-wide).

In July of 2016, American scientists proposed the Human Genome Writing Program (HGP-Write), with the goal of synthesizing a complete human within 10 years . The haploid genome consists of approximately 3 billion base pairs. The rapid development of synthetic biology, especially the start of the HGP-Write project, has made the demand for synthetic genes exponentially increase. A new generation of large-scale, low-cost, high-throughput gene synthesis and assembly technologies become a driving force in synthetic biology.

Synbio Technologies Synotype platform includes complete Syno® 1.0 – Syno® 3.0 DNA synthesis platforms. These three platforms have enabled us to form a complete DNA synthesis biological technology system, leading us in becoming an industry leader. As a reliable supplier of the new generation high-throughput chip DNA synthesis, we can parallelly synthesize tens to hundreds of thousands of primers onto a single semiconductor chip. Our innovative design and synthesis process and quality management ensure to providet the highest quality DNA fragments (including primers, elements, and genes), whole-gene synthesis, pathway synthesis, gene assembly, nucleotide pools, and various mutation library synthesis. Our genetic products and customized services cover the synthesis, construction, and shuttle of any DNA fragment into any vector system.

  • Our Syno® 1.0 – Syno® 3.0 complete DNA synthesis technology platforms are designed to meet our customers’ individual needs in terms of throughput, delivery cycle, price, difficulty, and so on.
  • Syno® 3.0 high-throughput DNA synthesis platform, with prices as low as $0.09/bp.
  • Ensure that the delivered gene sequence is 100% accurate.
  • Our patented NGTM Codon gene optimization software provides optimization for free and significantly improves protein expression levels.

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Synthetic DNA Variant Library Synthesis Service

Synthetic DNA library synthesis is widely applied mechanism within various fields of research such as: discovery biology, protein engineering, structural biology, and gene expression. All of these fields of scientific research have revolutionized the world we live in today, through various means. One of the more popular discoveries in the genetics research community was the construction of the first synthetic DNA library. A synthetic DNA library is a pool of DNA fragm ents which is stored within a host micro-organism via molecular cloning. The resulting synthetic DNA library encodes variants of a common design and structure. Each fragment is stored in a separate host organism, such as bacterium. This offers a palpable advantage due to bacteria’s ability to quickly and accurately replicate the DNA to produce detectable quantities. This process allows us to have quick access to many different sequences of interest with ease and efficiency.

Compared with traditional methods for DNA synthesis

The traditional method of “design-build-test” routine to reach a genetic design to improve synthetic biology has been proven to be slow, poor, and unreliable. Due to this reason, an alternative method of generating a DNA libraries was discovered and utilized within the research community with higher frequency. With the Synthetic DNA library synthesis service, high performing genetic solutions will be worked out with ease and efficiency. The DNA library synthesis pool can achieve gene synthesis and cloning with a much faster, smarter, and more reliable approach. This allows for a much more accurate and efficient DNA library to be synthesized with the use of DNA synthesis. With a more accurate and efficient DNA library, various topics within genetics research will better understood and more effectively studied.

Synthetic DNA library synthesis makes biological engineering much more efficient

With DNA library synthesis based methods, highly complex sequences within synthetic biology can be engineered rapidly with high accuracy. It is a novel approach which allows for the customer specific DNA sequence to be more reliably synthesized and obtained with ease and higher accuracy. This also enables new biological systems and products to move toward potential revolutionary discoveries.

Custom Synthetic DNA library synthesis services

DNA synthesis allows us to synthesize gene libraries of any size and complexity, specific to the customer’s specifications. DNA library synthesis relies upon DNA synthesis, a method mastered by Synbio Technologies through our three phase Syno® Platform. This platform allows us to generate the customer specified DNA sequence of interest, up to and including 150 Kb in length, with one hundred percent accuracy. DNA synthesis is the technology that provides the foundation to meet the needs of the DNA library construction and maintenance. This strong foundation is important because DNA synthesis is the first step for generating DNA libraries. With the proper construction of DNA libraries researchers aim to identify novel genes and how each gene relates to various protein functions and structures.

DNA Synthesis Related Services

  • Codon Optimization
  • gene synthesis
  • Small Genome Synthesis
  • Pathway Synthesis

Progress in DNA Synthesis Steps

Within the last few decades, advancements in methods of DNA synthesis, as well as synthesizing, DNA sequences have improved our capabilities of understanding genomics which has led to improved capability of engineering biological systems. The chemically synthesized DNA oligonucleotides and their assembly constructs contain encoded genes, synthons, circuits, metabolic pathways and even whole genomes. These assemblies have the ability to design, build, and test various research topics within molecular technology and synthetic biology. At Synbio Technologies we pride ourselves in our ability to successfully synthesize the requested DNA sequence specific to each customer. We have the ability, through our Syno Gene Synthesis Platform, to synthesize sequences up to and including 200kb in length with guaranteed one hundred percent accuracy.

DNA synthesis from oligonucleotides

As a polymer, consisting of four different phosphoramidite monomers, DNA synthesis follows a form of hierarchical polymer synthesis. Within the more traditional method of synthesizing DNA, nucleotide monomers make a connection between each other in order to construct individual oligonucleotide. On the other hand, in a more modern method of conducting gene synthesis, for assembling a synthetic double-stranded DNA, namely synthon, requires that adjacent oligonucleotides should be equipped with overlapping sequences. Once the sequences are long enough, the sequences-verified synthons are assembled to produce synthetic DNA constructs. The synthetic DNA constructs can then be commonly found within metabolic pathways or whole genomes.

Traditional column-based oligonucleotide as a source of DNA synthesis has a limitation to cost reduction. The cost of that ties to the consumption of reagent use, the scale of synthesis, the length of the oligonucleotides and the technical supplier.

DNA synthesis from high-fidelity microchips

The demand of cheaper resources necessary for DNA synthesis has resulted in an interest in a more recently developed method of array-synthesized DNA. Within the original method of DNA synthesis, the oligonucleotides were attached to a microchip surface. This attachment was accomplished via applying a modified phosphoramidite synthesis steps. This promising route has the capability to generate up to a million diverse oligonucleotides for constructing DNA on an individual chip.

Admittedly, oligonucleotides from DNA microchips can minimize the cost of DNA synthesis by a significant magnitude. High-fidelity microchips are able to be optimized through controlling depurination during the process of synthesis.

Synbio Technologies has in placed Syno® 3.0 synthetic platform, which is a next generation chip based DNA synthetic technology. Tens of thousands of or even one hundred thousand primers for one-time synthesis can be achieved on a semiconductor chip. Synbio Technologies is competent to provide lower-cost methods to produce high-quality synthetic DNA to meet all customers’ request.

DNA Synthesis Related Services

  • Codon Optimization
  • gene synthesis
  • Small Genome Synthesis
  • Pathway Synthesis

DNA Synthesis Definition and Methods

DNA synthesis definition

DNA synthesis is defined as the process by which copies of nucleic acids are strung together to form a longer DNA sequence within a laboratory setting.

DNA synthesis processes

DNA oligomers are the foundation of the DNA synthesis 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 150 Kb in length with one hundred percent accuracy, guaranteed to be identical to the sequence requested by the customer.

DNA synthesis assembly in yeast

A commonly used method of DNA fragment assembly is through the use of a yeast intermediate. The yeast, better known as Saccharomyces cerevisiae, can synthesize single-stranded DNA with a minimum length of 38 overlapping base pairs and a linear double-stranded vector in one transformation event. In this way, oligonucleotides can synthesize DNA for a wide range of lengths, ranging from as few as 20bps in length to as large as 200bps in length. DNA synthesis assembly in yeast is a useful method for building synthetic DNA molecules that are relatively large in length. Since yeast have a powerful capacity to synthesize and recombine DNA fragments, yeast has become a model eukaryote for studying numerous cellular processes. In addition to this advantage, homologous recombination in yeast can be used to build DNA fragments from overlapping constituent parts.

TopDown one-step DNA synthesis method

TopDown DNA synthesis is a simple and cost-effective method to assemble a long genetic sequences up to and including 150 Kb in length. The TopDown DNA synthesis method can be distinguished from conventional gene synthesis methods by two key features. First, the melting temperature of the outer primers is designed to be ~8°C lower than that of the assembly oligonucleotides. Secondly, there are different annealing temperatures that are utilized to selectively control the efficiencies of the oligonucleotide assembly and full-length template amplification. The TopDown DNA synthesis method is an effective way to remove the interference between PCR assembly and amplification. This interference between PCR assembly and amplification have been linked to the development of certain malfunctions within the DNA synthesis process. This demonstrates that TopDown DNA synthesis is a method which can optimize DNA synthesis under certain condition.

Synbio Technologies is a DNA technology company whose main focus is DNA synthesis. 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 150 Kb 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.

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Synbio Technologies’ DNA Synthesis Service

DNA synthesis is a method in synthetic biology for the creation of deoxyribonucleic acid biopolymers in a sequence-specific manner. DNA synthesis can be further classified into oligonucleotide synthesis and gene synthesis. Currently, oligonucleotides are manufactured almost exclusively using automated solid-phase methods.In this way, we can apply efficient and accurate DNA synthesis services. Oligonucleotide synthesis produces short, single-stranded nucleic acid polymers usually made up of 18 to 25 nucleotides, which are usually used as probes for detecting specific DNA/RNA sequences by recognizing and binding to a complementary segment of DNA/RNA. Oligonucleotides are widely used as DNA/RNA structure probes, DNA microarray probes, and gel electrophoresis references, and are usually single strand fragments limited to 100 nucleotides. On the other hand, gene synthesis is able to create double-stranded DNA ranging from 50 bp to 12,000 bp in length. Solid-phase based gene synthesis opens up new avenues for completely synthetic double-stranded DNA molecule with no apparent limits on either nucleotide sequence or size.

Synbio Tech DNA Synthesis Service

Taking advantage of industrial-leading processing technology, Synbio Technologies’ Syno® 1.0 platform provides a full selection of conventional primers, ultra-high purity primers, degenerate primers, aptamers, and fluorescent probes for world-class scientific research and some industrial customers.Synbio Technologies can also manufacture large scale primer synthesis using multi-well plates to deliver up to 10,000 primers for customized orders.

Our Syno® 2.0 DNA synthesis platform is the most versatile platform empowered by patented Syno® DNA Assembly technologies. Syno® 2.0 platform can cope with any type of gene and genome ranging from 100 base pairs to 1 million base pairs that contain all types of complex sequences. Standard complexity order can be completed within 5 business days with 100% sequence fidelity. NGTM software is developed at Synbio Technologies to boost the expression of synthetic genes in all commonly used expression systems.

Next Generation DNA Synthesis Company

As a leading DNA synthesis company, Synbio Technologies offers a comprehensive package of synthetic genomics services, from simple oligonucleotides and DNA fragments to large and complex genome clusters. Our cutting edge Syno® 3.0 high-throughput platform is the world’s most advanced next-generation DNA synthesis platform, able to provide affordable synthetic DNA at unprecedented scale.

At Synbio Technologies, our DNA synthesis capacity is empowered by several patented technologies. Industry-Leading NGTM Codon can help customers redesign and enhance the efficiency of the genes to achieve superior gene expression. Syno® Assembly is truly one-of-a-kind technology, working in tandem with our Syno® 2.0 platform to ensure the delivery of affordable, error-free sequences.

In addition, Synbio Technologies is building up its first integrated Syno GPS (Genotype, Phenotype and Synotype) system, aiming to streamline translation or reverse translation between “Genotype” and “Phenotype” through our proprietary “Synotype” platform. Our company’s scientific capabilities encompass areas such as molecular biology and bio-pharmaceutical research, precision medicine, biotechnology, molecular breeding, and biofuel production.

Synbio Technologies, a DNA synthesis company devote to next generation DNA synthesis, provides a variety of synthetic genomic options to advance your research, including primer synthesis, gene synthesis, metabolic pathway synthesis, gene cluster synthesis, CRISPR-Cas9 service, Sanger sequencing, and NGS services.

DNA Synthesis Application

With the growth of DNA synthesis techniques, especially the focus on automation and automatic technologies, we can more easily, rapidly, and efficiently synthesize DNA fragments. The applications of synthetic DNA encompass a broad spectrum ranging from genetic circuits and metabolic pathways to synthetic genomes. DNA synthesis is an essential technique in molecular biology and plays a vital role in various disciplines, such as genetic engineering, and clinical diagnosis/treatment.

DNA Synthesis in Molecular Biology

According to the clarified nucleotide sequence and amino acid sequence, we can synthesis the polypeptide and protein gene which can be applied in actual production and research. So far, various genes have been synthetic and expressed successfully in eukaryotic system, such as human growth hormone, interferon, insulin, interleukin etc.

DNA Synthesis Applications in Medicine

As our technology has become more powerful, our understanding of many diseases has become more sophisticated, from the organ and tissue level to the molecular and genetic level. As a result, new discoveries have allowed us to tackle genetic diseases in ways that we could not previously. For example, fluorescent probes can detect potentially harmful deletions in certain genes, proving to be a valuable tool for clinical diagnosis.

DNA Synthesis Application in Synthetic Biology

A valuable domain of synthetic biology is metabolic engineering. By engineering parallel metabolic systems that interface with natural cellular metabolic machinery, researchers can program cells for practical applications including the synthesis of cost effective chemicals or drugs. In one demonstration by Keasling and colleagues, the construction of a biosynthetic artemisinin pathway in yeast enabled the microbial production of this antimalarial compound at one-tenth of the cost of conventional production method of harvesting from the rare Artemisia annua plant.

Synbio Technologies concentrates primarily on DNA synthesis technologies. We have established our proprietary Syno® gene synthetic platform which can provide a variety of services including oligonucleotide synthesis, fluorescent probe labeling, and synthesize gene/genome, pathway and DNA library. We can also provide materials and supplies for clients in various disciplines, such as biological medicine, molecular diagnosis, industrial enzyme, bio-fuel, modern agriculture, environmental protection and surveillance and DNA storage.

Gene Synthesis Services

At Synbio Technologies we offer a unique approach to gene synthesis services through our revolutionary Syno Gene Synthesis Platform. This three phase platform allows us to generate any customer specified genetic sequence in text format, up to and including 150 Kb in length. The requested sequences can vary greatly in both length and structure. For example, a requested sequence can contain a gene with a deleterious mutation in order to induce a debilitating effect, rendering the gene inactive in order to study the phenotypic effects. Or the requested sequence can be one not previously existing within nature, in order to study the effects of evolution or antibiotic resistance. These examples are made possible through Synbio Technology’s various gene synthesis services. First, our Syno® 2.0 Gene Synthesis plays a large role in small gene/short sequence synthesis. The Syno® 2.0 Gene Synthesis platform may be meant for short sequence synthesis, but it still has the capabilities and confidence of generating sequences with extreme accuracy of up to and including 150 Kb in length. Second, our Syno® 3.0 Next Generation Gene Synthesis Platform is a groundbreaking piece of technology that allows us to synthesize sequences that span near the length of small genomes. This gene synthesis services allows us to generate sequences up to an including 150 Kb in length, which was previously thought to be impossible to successfully and accurately engineer in a laboratory setting. With these two gene synthesis services, along with a wide range of other biological technology services, Synbio Technology is ready to supply our customers with the highest quality product within an efficient timeframe. This confidence starts with one of our most successfully gene synthesis services, the Syno® 2.0 Platform.

The Syno® 2.0 Gene Synthesis Platform is one of the corner stones of our gene synthesis services. This platform has various specializations in gene synthesis such as: pathway synthesis, small genome synthesis, vector construction, and codon optimization technology. With this platform, we are able to successfully and accurately synthesize the customer specified sequence of interest with one hundred percent accuracy. This is accuracy and confidence in sequence validity comes from the multiple occasions in which we use Sanger sequencing to verify the sequence being generated. In addition to the high quality sequence synthesis, we offer cloning of each prepared sequence to ensure the customer is provided with the adequate amount of product. Before each sequence is amplified, it is verified again with Sanger sequencing to verify the accuracy of the sequence. In addition to the specializations Syno® 2.0 offers, this platform also offers the ability to generate various structures of DNA commonly utilized in various fields of genetics research. Some of these structures include: hairpin structures, repeated sequences, high GC content regions, high AT content regions, or multiple consecutive nucleotide sequences. All of these structures offer a unique approach to studying various topics within the field of genetics. With the Syno® 2.0 Platform, and our other gene synthesis services, Synbio Technologies is ready to offer our customers a unique approach to conducting genetic research.

The Syno® 3.0 Next Generation Gene Synthesis Platform differs slightly from the Syno® 2.0 Platform, and our other gene synthesis services, but the output is still guaranteed to be one hundred percent accurate with a low price and fast turnaround time. The Syno® 3.0 Platform specializes in ChipBased DNA synthesis, CRISPR-Cas9 Libraries, and digital information storage. In other words, the Syno® 3.0 Platform is one of our gene synthesis services that offer a large amount of generated sequences to be synthesized. The chip-based gene synthesis approach allows us to generate thousands upon thousands of sequences all within a single chip. This allows for rapid production of sequences with high accuracy when multiple chips are used. The generated sequences are then, of course, verified with Sanger sequencing to guarantee one hundred percent accuracy. This method is used to generate large genes, up to and including 150 Kb in length with one hundred percent accuracy. In addition to this, the Syno® 3.0 platform gene synthesis services that Synbio Technologies offers, is the use of and experience with CRISPR-Cas9. Ever since the discovery and first successful utilization of CRISPR-Cas9, in 1993, it has been used time and time again in order to conduct genome edits. CRISPR-Cas9 has become one of the most popular and effective methods in order to conduct genome edits. This gene synthesis service offers CRISPR-Cas9 based gene knockout-out, gene knock-in, transcription activation, and transcription inhibition. All of these applications are methods of altering the expression level of a particular sequence/gene of interest. This alteration of expression will then be applied to various types of genetics research.

All of the gene synthesis services, as much as they vary in application, offered by Synbio Technologies have been tested and proven to be one hundred percent effective. This comes in the form of our guaranteed sequence validation, through Sanger sequencing. This accuracy and validity have given Synbio Technologies a reputation as one of the premier companies in the gene synthesis industries. This reputation relies heavily on our Syno® Gene Synthesis Platform. With this gene synthesis service platform, we are able to synthesize the sequence/gene of interest with extreme accuracy in a cost effective manner. This cost effective manner, on top of the efficient turnaround time, we at Synbio Technologies are ready to supply our customers with the highest quality product for the lowest prices.

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