Many consider the twentieth century the "century of the atom" due to the many innovations made possible by new understandings of the elements' physical and chemical properties. These included, but are not limited to, the development of atomic energy (and weapons), diagnostics for medicine, computers, and the microchip, among others.

These breakthroughs shaped the potential and future of science and technology and profoundly impacted our daily lives. But we're living through what may be dubbed the "century of DNA." DNA is like the soundtrack to life's complex symphony; it serves as a template that dictates how cells work. Recent decades have seen tremendous progress in reading and synthesizing DNA sequences, greatly enhancing our capacity to understand and manipulate biological systems.

In today's post, we will consider two major branches of biological engineering: gene synthesis and traditional cloning. This blog will review the main benefits and drawbacks of each method to assist you in choosing the appropriate media for your project.

Let's begin with a closer look at the two camps.

What is Gene synthesis?

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Gene synthesis is the in vitro method of producing a gene without requiring a preliminary template.

Alternatively, if you would want a more in-depth explanation, synthetic gene creation is a technique that involves a series of steps for producing a gene and other gene derivatives without the need for a DNA template. As a result, it eases the process of making various genes, notably custom genes with unique base pairs and sequences.

Har Gobind Khrona and colleagues performed the first effective gene synthesis using a yeast tRNA in 1972. Since 1972, advances in gene assembly andmolecular biology techniques have allowed the successful extraction of genetic sequences unique to a broad range of scientific issues.

Steps Involved in Gene Synthesis

A general overview of the steps towards a successful gene synthesis include:

• Oligonucleotide synthesis

• Oligonucleotide annealing
• The cloning of gene sequences
• Screening for clones
• Analyzing sequences and fixing mistakes

A couple of decades ago, the idea of using gene synthesis for targeted gene therapy or other intriguing subjects would have appeared entirely out of reach. Gene synthesis opens up a world of possibilities beyond what has been explored so far, with applications spanning from gene regulation to a greater understanding of evolutionary development and antibiotic resistance, among many others.

Thanks togenetic engineering, we can now construct custom antibodies, variant libraries, custom peptides, operons, and enhanced protein functions.

Now, let's turn our gaze to the more traditional method of doing things.

What is Traditional Cloning?

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Traditional cloning is a type of DNA manipulation that typically involves employing restriction endonucleases to produce DNA fragments possessingparticular complementary termini, which are subsequently ligated together with a DNA ligase before undergoing transformation.

To do this, you usually need to cut the DNA sequence in two places so that there are two restriction enzymes on either side of the DNA sequence. These enzymes also reside at the best place for the vector to insert itself, sometimes called the multiple cloning site (MCS).

By applying two separate REs, incompatible ends are produced, leading to the directional cloning of the insert and reducing the transformation background of the re-ligated vector only.

Pros and Cons of Traditional Cloning

Pros

• Low costs

• Flexible
• A wide variety of vector options
• Performing directional replication is easy and stress-free

Cons

• A restriction site's existence and translation may impose sequence limitations.

The first step in conventional cloning using recombinant DNA technology is to use restriction enzymes to prepare a vector to accept an insert DNA. Following digestion, the fragments are joined by the enzyme ligase, a procedure known as ligation, creating a new vector that can express a desired gene.

This is the most basic and old-fashioned method of genetic material production, and it served as the basis for the development of novel cloning techniques that capitalize on the unique properties of other modifying enzymes, including:

• TA cloning
• TOPO cloning
• PCR cloning,
• Ligation-independent cloning, and
• gene assembly.

You can simplify the steps described above for typical traditional cloning as:

• Vector preparation
• Insert preparation
• Ligation Transformation
• Colony screening

Given this information, let's contrast the two (gene synthesis vs traditional cloning) to better understand their differences.

Traditional Cloning vs. Gene Synthesis: Major Differences

When compared to molecular cloning and polymerase chain reaction (PCR), two of the more conventional approaches to gene cloning, gene synthesis has numerous significant benefits. There are two main limitations to these two traditional approaches. First, they can only amplify and clone genes; they cannot synthesize them.

Another limitation of restriction molecular cloning and polymerase chain reaction is that the genetic code of interest must first be acquired and then amplified. This is a prerequisite for both techniques. With gene synthesis technology, the desired genetic sequence can be amplified without physically obtaining it, which is a significant upside.

Current gene expression technologies enable the synthesis of "de novo" sequences. This allows for synthesizing sequences of interest even without physical copies of genes, such as viral DNA or cancer cell mutations in particular genes. The lack of a physical replica of the target gene considerably reduces the financial and time investments required in contrast to conventional approaches such as cloning, amplification, gene target knockouts, and targeted mutagenesis.

Owing to this benefit, gene synthesis has grown in popularity across several scientific domains.

Therefore, gene synthesis has been utilized extensively in numerous scientific domains due to its user-friendliness and cost-effective procedure.

Suffice it to say both techniques have unique applications based on the requirements of a given project.

Continue reading to learn some essential applications of these two approaches.

Applications of Gene Synthesis and Traditional Cloning

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The Uses of Gene Synthesis in Real-Life

1. Vaccine Research And Development

In vaccine research, gene synthesis is widely employed in everything from fundamental immunology and microbiology investigations essential for logical vaccine design to state-of-the-artDNA synthesis and vaccine development initiatives. The development of new vaccines is necessary to combat both newly emergent dangers, such as the Ebola virus, and persistent infectious viruses, such as the seasonal influenza A virus.

Meanwhile, cancer vaccines have twofold purposes: first, as a preventative measure against oncogenic virus infection; second, as a therapeutic tool to mount an immune response against malignancies.

Building codon-optimized custom genes encoding adjuvants, customized vectors tailored for safety, and gene cassettes that control the expression of specific antigenic proteins are all possible using gene synthesis.

2. Plant Biology Research

Gene synthesis can be deployed for developing DNA vaccines by creating gene cassettes that make specific antigenic proteins, codon-optimized synthetic genes that make adjuvants, and safety-optimized special vectors.

3. Therapeutic Antibodies

It is possible to activate, suppress, or modify endogenous immune responses to particular cells or molecules with therapeutic antibodies, particularly monoclonal antibodies. The treatment of numerous diseases, including cancer, inflammatory and autoimmune disorders, and many others, is being transformed by antibody-based medications. Some examples include:

• Therapeutic Inhibition of Inflammation in Pigs and Humans by Chimeric Anti-CD14 IGG2/4 Hybrid Antibodies.
• In experimental autoimmune encephalomyelitis in mice, removing autoantibodies improves the symptoms.
• One Human Monoclonal Antibod y Cross-Neutralizes Four Paramyxoviruses.

4. Brain research

A lot of neuroscience research uses gene synthesis. For example, codon-optimized channelrhodopsins make it possible to use optogenetic methods to see how neurons are connected; customized ORF clones and RNAi constructs help to understand how glioblastoma cells communicate; and gene variants are used to look into how neurons communicate and change how they connect and learn, to name a few examples.

The Uses of Traditional Cloning in Real-Life

• The traditional cloning of genes empowered the development of proteins, medicines, and other products. These can take many forms, including cloned DNA, cells, tissues, or an animal.
• When it comes to agriculture, cloning is utilized to develop pest-resistant vegetation.
• In addition, cloning is exploited for gene therapy and the creation of transgenic animals.
• The process of cloning cattle allows for duplicating desirable traits, such as fast growth and high milk output, absent the genetic "lottery" and endless blending during sexual reproduction.
• It allows for the faster breeding of an animal with a specific genetic modification compared to regular mating, for example, the ability to produce medication in milk.
• Furthermore, modifying a gene in cultured cells is much simpler than doing so in an entire animal. Moreover, combining the altered cell nucleus with an enucleated egg makes a clone of the desired genotype possible.
• Animals used in experiments, like mice, are cloned so researchers may learn more about their fundamental biological processes.

Choosing Between Gene Synthesis Services And Traditional Cloning Services

Although numerouscustom gene production services exist in the market, the right one usually offers supplementary benefits that extend beyond the DNA itself:

Here are some things you should look for when choosing Gene synthesis services.

Dependability: Over 21 years of experience in the synthesis of synthetic DNA and 5-star Google review; dependable I00% sequence-verified genes assured by ISO-accredited quality systems; and environmentally responsible production practices.

Enhanced efficiency and economy of scale: With the right exclusive program, you can improve your sequence for dependable and enhanced expression in your host system while reducing the cost per milligram of protein.

Stellar Customer Experience: Placing an order for synthetic biology products has never been easier. It's now possible to request a quote at any given moment and location at the click of a button. You won't have to wait days for an estimate or optimization outcomes. The only thing you may need to do is submit your sequence, choose your criteria, and then place your order. Any and all questions you may have about gene synthesis can be quickly addressed by contacting our knowledgeable customer service team.

What you require as a product: Depending on your requirements and budget, you may decide between a variety of custom genes, antibodies, or custom peptides from our product lineup. Or you can choose to create your project from the ground up. We offer a range of products, from DNA fragments to expression-ready designs, DNA libraries, and protein expression and purification services. Everything is handled internally by our experts.

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High Success Rate: Our proficiency incustom gene synthesisis substantiated by the fact we excel in producing genes with multiple complexities whileaccomplishing exceedingly challenging sequences with a success rate of 99%.

Variety of Platforms: Companies that exclusively depend on silicon-based platforms are limited to developing simple and short genes, which can hinder their capacity to work on a significant portion of gene projects—up to 50%.

Our accomplishments may be attributed to more than two decades of experience, during which we have created and employed special reagents and enzymes in our manufacturing procedures, all while following the most reliable synthesis techniques.

Summary

Commercial gene synthesis services are rapidly becoming a viable, less expensive substitute for conventional molecular biology methods like cloning, which goes against the grain of what many researchers previously believed. The primary reasons are as follows:

• Reduced workload - The conventional approach to cloning is a multi-step procedure that comprises the design of a cloning strategy, primer production, polymerase chain reaction (PCR), gel sampling, and bacteria transformation, among other sophisticated steps. Unlike gene synthesis, these procedures demand a substantial investment of time and human resources.
• Minimizes expenses - Most of the time, ordering a manufactured gene is cheaper than ordering cloning kits, oligos, or DNA/gene sequencing services.

• Improved DNA performance -  Codon optimization is possible during gene synthesis and has been shown to make protein translation more efficient.
• Practicality - Researchers can get any gene they want by just providing the nucleotide or amino acid sequence. This is because they don't need a physical frame and aren't limited by design constraints that come with traditional cloning.

Irrespective of which camp (gene synthesis or traditional cloning) you decide to join, you'll need reliable, high-quality supplies like Elisa kits, bioreagents, and peptides. Luckily, Biomatik is a one-stop shop for these biochemical products, where you can find services like custom gene synthesis, recombinant protein production, and more. Reach out today!