Category: PCR Cloning

Molecular Biology Evolution

Since molecular biology was first established, in the 1930s, the complexity of various biological systems have been explored in order to better understand these systems. As years went by, and our understanding of these systems increased, the information that fell under the, large and ever-growing, umbrella of “molecular biology” became more specific and eventually subdivided by the particular sub-field being studied. Some of these sub-fields include: genetics, proteomics, and cytology. Many of these sub-fields have become extremely popular and very well studied since molecular biology was first established.

Genome Project and Molecular Biology

In the more recent past, there has been an influx of quantitative research within molecular biology. This groundbreaking research has completed the connection between two popular sub-fields of molecular biology: computer science through use of bioinformatics and computational biology. This was most notably seen with the success of the Human Genome Project (HGP). The Human Genome Project allowed for over three billion nucleotide base pairs of euchromatic genome to be sequenced and presented as a reference. This was an important milestone of molecular biology which allowed the human genome to go from the analog world of biology into the digital world. This was a giant step, within the field of molecular biology, which caused a domino effect that resulted in thousands of human genomes to be sequenced with increasingly lower cost.

On 2rd June 2016, the Genome Project-Write (GP-write) announced a continuation of genomics research through multiple molecular biology tools. These multiple molecular biology tools included gene synthesis and genome editing technologies. These technologies are utilized to synthesize and test large portions of many genomes stemming from microbes, plants and even animals. GP-write’s commitment to furthering our understanding and effectiveness of these technologies has the possibility to improve research and development of the topics of life sciences, new bio-based therapies, and nutrition.

New era of molecular biology

Even as these recent accomplishments in molecular biology unfold, a new problem seems to be stemming from them. The problem that has come about comes in the form of designing and even artificially synthesizing new life.

A recent study has proven the ability to accomplish a complete chromosomal transplantation from one cell to another. After the transplant has been successfully conducted, the chromosome can then be activated to conduct various genetic activity. We can then utilize specific enzymes, digestive proteins and other substances within these cells. This combination will result in the cell’s loss of original features and a totally new species.

Due to the crisis of resource shortage critical to human sustainability partnered with the ever-increasing human population, there is a need for us to seek effective approaches for sustainable living. The furthering of our knowledge on biological systems through these technologies would have many positive effects on successfully creating a sustainable habitat. These positive effects would come as a result of better understanding of the physiology of cells, developing new molecular medicines, as well as generating sustainable energy sources, such as biofuels. All of these will contribute to a more successful living environment through the use of molecular biology.

One molecular biology technology, gene synthesis, has slowly become better understood over the past ten years and has drastically increased our capability of editing and synthesizing genes of interest. Synthesizing DNA artificially is very difficult and increases in difficulty when attempting to synthesize long genetic sequences. This is because the longer the sequence being generated is, the higher the possibility of generating errors. Therefore, a new method is required to successfully conduct gene synthesis and correct for all mistakes generated when the sequence was being synthesized.

As one of the leading companies in the field of synthetic biology, Synbio Technologies has unique proprietary GPS platform on the basis of genotype, phenotype and synotype. We can provide excellent molecular biology services including: plasmid DNA preparation, PCR cloning and subcloning, site-directed mutagenesis and vector construction. We have the ability to generate sequences that are de novo, meaning that the genetic sequence is not preexisting within any organism in biology. We also have the ability to generate sequences up to and including 200kb in length in addition to complex gene products and structures. Synbio Technologies prides itself in our ability to use these molecular biology technologies to better suite our customer’s needs when conducting various types of research.

Molecular Biology Related Services

Gene Synthesis and Cloning

Gene Synthesis and Cloning is an integral part of many research pipelines. There are two popular methods of genetic cloning: DNA amplification by use of subcloning and polymerase chain reaction (PCR). The former needs more time and resource consuming, while the latter spends less time and cost effectively. Traditionally, subcloning was commonly used for amplification. But recently PCR has become much more effective and popular. The popularity of amplification by subcloning may have dwindled because it requires multiple steps. The process starts with the removal of the genetic sequence of interest using specific restriction enzymes. This same restriction enzyme is then used to open up the plasmid where the genetic sequence will be inserted into. This creates “sticky ends” which allow the genetic sequence to be more easily inserted. The genetic sequences is then added into a plasmid and sealed in place with DNA ligase. The plasmid is subcequently inserted into bacteria where the genetic sequence of interest is amplified and later extracted. This is a somewhat laborious process when compared to PCR. PCR has very simple steps: extracting the genetic sequence of interest and loading it into the machine. The sequence of interest is then rapidly amplified with a high quality output. The amplified sequence can then be used to in various research topics specific to the user. Amplification of these sequences were once restricted to only allow preexisting sequences to be extracted and amplified, but recent technology has allowed a small change in this.

The interesting aspect that Synbio Technologies offers is the ability to synthesize the genetic sequence of interest, as opposed to extracting the preexisting sequence from an organism. This technology is referred to as our Syno®2.0 Platform, which allows Synbio Technologies to successfully synthesize any desired genetic sequence, specified by the user. The lengths of these sequences can vary from a small as a few hundred base pairs to up to 100kb or even more in length. The Syno®2.0 Platform provides an extremely effective and revolutionary approach to genetic research. Traditionally, the sequence of interest must be present within a genome of an organism to then be extracted and later amplified. This platform allows us to synthetically create a sequence that does not need to be preexisting within any organism. The Syno®2.0 Platform allows us to move from a text file of the user specified sequence to the physical genetic constructs, bypassing the previous stipulation of extracting the preexisting sequence. The resulting synthetic products is then analyzed using Sanger Sequencing in order to verify and guarantee one hundred percent accuracy. Once successful, Synbio Technologies offers the option to clone this sequence as many times as necessary. This is done by use of either PCR or subcloning, both of which are extremely effective mechanisms to amplify the genetic sequence. The process of gene synthesis and later gene cloning allows Synbio Technologies to supply the researcher with a sufficient amount of genetic material needed within an efficient timeframe.

Synbio Technologies also offers PCR cloning if the requested sequence of interest is already present and does not need to be synthetically engineered. First the provided sequence will be verified using Sanger Sequencing in order to account for one hundred percent accuracy. The resulting verified sequence will then be put through our flexible and reliable pipeline of gene cloning. Using patent pending, Syno®2.0 Platform and Clone®3.1 system, Synbio Technologies is able to achieve inserting any sequence of interest into any site of a vector specified by the user. This allows the user to specify which restriction enzyme to use, both for the extraction of genetic sequence, as well as the restriction enzyme used on the plasmid. These wide range of possibilities allows the researcher to design a location within a specific vector in order to amplify the sequence of interest. This specificity is something that Synbio Technologies is very proud of, allowing us to adhere to any requests that the user might have while also providing a high quality output in the process.

At Synbio Technologies we pride ourselves as being one of the leading companies in the biotechnology industry, especially when referring to gene synthesis. We offer both accurate and time effective approaches to gene synthesis and gene cloning. The genetic sequence of interest will be Sanger Sequenced and verified for both genetically synthesized sequences as well as sequences provided by the user. This allows us to verify that the sequence, as well as the resulting amplified sequences, are extremely accurate before shipping the resulting product to the user. If the DNA sequence of interest has not yet been sequenced with high quality, we will conduct sequence validation in order to verify the accuracy before amplification. In addition to the accuracy and time efficiency, Synbio Technologies offers competitive prices for both gene synthesis and cloning of your sequence of interest. With competitive prices, along with the verified accuracy and time efficiency, Synbio Technologies is ready to offer customers a unique pipeline to utilize and rely on when conducting various types of genetic research that including gene synthesis and gene cloning.

Applications of PCR Cloning Technology

PCR cloning technology can amplify trace amounts of DNA, generating millions of copies of a specific DNA sequence. PCR is highly sensitive, extremely specific, and high-yield, while also being easily reproducible, fast, and convenient, making it an amazingly powerful tool in molecular biology. With the rapid development of modern life science, PCR cloning technology has been more and more widely applied to various fields in biological research, medical research, virus detection, and the food industry.

PCR cloning Technology Applied to Gene Cloning

Gene cloning and Subcloning via PCR technology plays a significant role in cell biology research. PCR technology can generate millions of copies of a single-copy gene, amplifying a specific DNA fragment that might only be a few picograms. Compared to other gene cloning techniques, PCR omits several tedious processes for cloning of a particular gene fragment from genome DNA, such as enzyme digestion, connection, transformation, DNA library construction, gene screening, gene identification, and Subcloning.

PCR Cloning Technology Applied to DNA Recombination

In molecular biology, PCR cloning technology can be used to construct recombinant DNA molecules by inserting different sources of specific genes or DNA fragments into viruses, plasmids or other vectors in vitro. Recombinant DNA molecules are then imported into reporter cells to amplify and reproduce. After screening, the daughter cells that contain the target gene are further multiply to extract a large amount of DNA. Recombinant DNA technology can be applied to the Human Gene Project, valuable protein expression, gene diagnosis and therapy, genetic modification of animals and plants, and other research fields.

PCR Cloning Technology for Gene Quantification

PCR cloning technology can be applied to quantitatively determine the copy number of a target gene in a sample. The target gene and a single copy reference gene are placed in a tube for PCR cloning. The PCR product is then separated by electrophoretic separation technology and band intensity is observed. Alternatively, the 5’ end of the primer can be marked by a radionucleotide, after which the gene copy number can be determined by radioactivity quantification. PCR cloning technology can also be applied to quantitatively analyze mRNA and tRNA. It can even detect 1TIRNA, which is hard to detect even by Northern blot.

Alteration of endogenous genes and invasion of foreign genes can be threatening to human health. Regardless of whether or not pathogenesis is caused by genetic changes, as long as there is a pathogen, its corresponding existing nucleic acid can be found. With the development of PCR cloning technology and related technologies, PCR can be applied to infectious disease pathogenesis detection and diagnosis, tumor related gene detection, hereditary disease early diagnosis, bone marrow transplant HLA – D locus matches, and evolutionary theory analysis.

Synbio Technologies’s Syno®2.0 platform can clone a target gene to any specific point on provided vectors without depending on restriction enzyme sites, and can quickly and cost-effectively fulfill a wide variety of client requests for myriad applications in synthetic biology.