Download PDF
Download XML
DNA sequencing is the intricate process of determining the exact sequence of nucleotides—adenine (A), thymine (T), cytosine (C), and guanine (G)—that compose a DNA molecule. This precise order of nucleotides forms the blueprint for an organism’s genetic information. The significance of this process cannot be overstated; it has profoundly impacted numerous scientific and medical fields. In molecular biology, DNA sequencing has unveiled the complexities of genetic structures, allowing scientists to explore gene functions and regulatory mechanisms in unprecedented detail. Evolutionary biology has also been transformed, as sequencing enables researchers to trace the lineage and evolutionary history of species, providing insights into how life has evolved over millions of years.1-4
The completion of the first human genome sequence marked a monumental achievement in science. This monumental endeavor, known as the Human Genome Project, began in 1990 and culminated in 2003, costing approximately $2.7 billion. It involved an international collaboration of scientists and took about 13 years to decipher the sequence of about 3 billion base pairs that make up human DNA. The implications of this achievement were vast, offering a foundational reference for genetic research and heralding a new era in genomics.5-7
Since then, technological advancements in DNA sequencing have been nothing short of revolutionary. The advent of next-generation sequencing (NGS) technologies has dramatically decreased both the time and cost required to sequence a genome. These advanced methods enable parallel processing of multiple DNA fragments, vastly increasing throughput and accuracy. As a result, the cost of sequencing a human genome has plummeted from billions of dollars to just a few hundred dollars, making genomic information more accessible for research and clinical applications.8-10
Moreover, these technological leaps have enhanced the speed of sequencing. What once took years can now be accomplished in mere days or even hours. This rapid turnaround is crucial for applications such as personalized medicine, where timely genetic information can guide treatment decisions. In research settings, faster sequencing accelerates the pace of discovery, allowing scientists to quickly test hypotheses and explore new areas of genetic research.8,11
The broader implications of these advancements are profound. In medicine, they pave the way for more precise diagnostics and targeted therapies. In agriculture, they enable the development of crops with improved traits. In environmental science, they facilitate the monitoring of biodiversity and the detection of pathogens. Thus, the evolution of DNA sequencing technology continues to drive innovation across a spectrum of disciplines, opening new frontiers in our understanding of life and disease.12,13
The journey of DNA sequencing began with the groundbreaking work of Fredrick Sanger in 1977, a pivotal moment in the history of molecular biology. Sanger developed a method known as the "chain termination method," which revolutionized the field by enabling scientists to determine the sequence of nucleotides in DNA with unprecedented precision. This technique, often referred to as Sanger sequencing, was foundational in the development of modern genomics.
Sanger's method relies on several key components: a single-stranded DNA template, a DNA primer, DNA polymerase, normal deoxynucleotide triphosphates (dNTPs), and modified di-deoxynucleotide triphosphates (ddNTPs). The innovation of using ddNTPs, which lack the 3'-OH group necessary for forming a phosphodiester bond, was crucial. When a ddNTP is incorporated into a growing DNA strand, it prevents further elongation, effectively terminating the chain. This property allows for the generation of DNA fragments of varying lengths, each ending at a different nucleotide.
To sequence DNA using Sanger's method, the DNA sample is first divided into four separate reactions. Each reaction contains all four standard deoxynucleotides (dATP, dGTP, dCTP, and dTTP) and one of the four dideoxynucleotides (ddATP, ddGTP, ddCTP, or ddTTP). As the DNA polymerase synthesizes new DNA strands, the incorporation of a ddNTP halts the extension at specific points, resulting in a collection of DNA fragments of different lengths, each ending with a labeled ddNTP.
The next step involves separating these fragments by size using gel electrophoresis, a technique that allows the DNA fragments to be sorted based on their length. The separated fragments are then visualized using autoradiography or UV light, thanks to the radiolabeled or fluorescently labeled ddNTPs. The resulting pattern of bands on the gel provides a visual readout of the DNA sequence, which can be directly interpreted by reading the bands from the smallest to the largest fragment.
Advantages:
Low Error Rate: One of the most significant advantages of Sanger sequencing is its low error rate. The method's accuracy makes it a gold standard for DNA sequencing, especially for applications requiring high precision, such as clinical diagnostics and validation of sequences obtained by other methods.
Long Read Length: Sanger sequencing is capable of producing long read lengths, typically around 750 base pairs. This long read length is beneficial for sequencing complex regions of the genome and for assembling contiguous sequences with fewer gaps.
Ability to Target Specific Primers: The method allows for the use of specific primers to target particular regions of the DNA, making it highly versatile and useful for targeted sequencing projects, such as gene-specific studies or the analysis of specific mutations.
Disadvantages:
High Cost per Base: Despite its accuracy, Sanger sequencing is relatively expensive on a per-base basis. The cost includes reagents, radioactive or fluorescent labels, and the labor-intensive nature of the process, making it less suitable for large-scale sequencing projects.
Long Time Required to Generate Data: The method is time-consuming, requiring several steps, including multiple rounds of reaction preparation, gel electrophoresis, and data analysis. This extended timeline limits its throughput compared to more modern sequencing technologies.
Need for Cloning: Traditional Sanger sequencing often requires the DNA fragments to be cloned into bacterial vectors, a step that adds to the complexity and time required for the process. Cloning also introduces potential biases and errors, further complicating the sequencing workflow.
Overall, while Sanger sequencing laid the groundwork for the field of genomics and remains a reliable and precise method, its limitations in cost, speed, and scalability have led to the development of newer, high-throughput sequencing technologies that are better suited for large-scale genomic studies. However, its high accuracy and long read lengths continue to make it valuable for specific applications where precision is paramount.
Maxam-Gilbert sequencing, developed by Allan Maxam and Walter Gilbert in the late 1970s, was one of the first DNA sequencing methods. This technique involves the chemical modification of DNA followed by cleavage at specific bases. The process begins with the radioactive labeling of one end of the DNA fragment to be sequenced, typically by a kinase reaction using gamma-32P ATP. This labeling is essential for detecting the fragments after chemical treatment.
The method relies on chemical treatments to break the DNA at specific nucleotides. For instance, formic acid is used to depurinate purines (adenine and guanine), while dimethyl sulfate methylates guanine residues. Hydrazine is employed to modify pyrimidines (cytosine and thymine), and the presence of salt in the reaction can selectively inhibit thymine modification, allowing for a distinction between cytosine and thymine. After chemical modification, the DNA is cleaved at the sites of modification using a hot piperidine treatment.
The resulting DNA fragments are then separated by size using denaturing acrylamide gel electrophoresis. The radiolabeled fragments are visualized by exposing the gel to X-ray film, producing an autoradiograph. Each band on the film represents a DNA fragment of a specific length, allowing the sequence to be read from the pattern of bands.
Despite its initial success, Maxam-Gilbert sequencing fell out of favor for several reasons. The method is complex and labor-intensive, requiring multiple steps and the handling of hazardous chemicals. The use of large amounts of radioactive material poses significant safety and environmental risks. Additionally, the chemicals used, such as hydrazine, are highly toxic and pose health hazards to researchers. These drawbacks led to the development and adoption of alternative sequencing methods that are safer, more efficient, and easier to use.
Shotgun sequencing is a method that involves randomly breaking DNA into numerous small segments, which are then sequenced individually. This technique was a crucial step forward in the ability to sequence entire genomes. The process starts by fragmenting the DNA into small pieces, often using mechanical shearing or enzymatic digestion. Each fragment is then sequenced independently using the chain termination method (Sanger sequencing) or other sequencing techniques.
After sequencing, the individual reads are assembled into a continuous sequence using computer algorithms that identify overlaps between the fragments. These overlaps are used to reconstruct the original DNA sequence. Shotgun sequencing was instrumental in enabling whole-genome sequencing projects, including the Human Genome Project, as it allows for the sequencing of large and complex genomes by breaking them into manageable pieces.
Next-Generation Sequencing (NGS) Technologies
The advent of next-generation sequencing (NGS) has dramatically accelerated DNA sequencing capabilities, allowing for massively parallel processing and high-throughput sequencing. NGS technologies have revolutionized genomics by enabling the sequencing of millions of DNA fragments simultaneously, significantly reducing the time and cost associated with sequencing.
SOLiD (Sequencing by Oligonucleotide Ligation and Detection) technology, developed by Life Technologies, utilizes 2-base encoding to decode raw data into sequence information. This method generates a vast amount of small sequence reads, which are analyzed through multiple cycles of ligation, detection, and cleavage. Each ligation cycle involves the hybridization of a fluorescently labeled oligonucleotide probe to the DNA, followed by ligation and detection of the fluorescent signal. The process is repeated with different probes to build the sequence.
Advantages:
High Data Output: SOLiD sequencing can generate an enormous amount of data, making it suitable for large-scale genomic studies.
Multiple Cycles Increase Specificity: The use of multiple cycles of ligation and detection enhances the accuracy and specificity of the sequencing results.
Disadvantages:
Short Read Lengths: The read lengths generated by SOLiD sequencing are relatively short, limiting its application in certain genomic analyses that require longer reads for accurate assembly and interpretation.
Illumina sequencing, developed by Solexa (later acquired by Illumina), employs a sequencing-by-synthesis approach with reversible dye terminators. This method involves fragmenting the DNA and attaching adapters to the fragments. The fragments are then bound to a flow cell and amplified through bridge amplification, forming clusters of identical sequences. Sequencing occurs by adding fluorescently labeled nucleotides, one at a time, to the growing DNA strand. The fluorescent signal emitted by each incorporated nucleotide is detected and recorded, allowing the sequence to be determined.
Advantages:
Low Error Rate: Illumina sequencing has a low error rate, making it highly accurate for a wide range of applications.
Lowest Cost per Base: The cost per base of sequencing with Illumina technology is among the lowest, making it economically feasible for large-scale projects.
High Data Throughput: Illumina platforms can generate a vast amount of data in a single run, enabling the sequencing of entire genomes quickly and efficiently.
Disadvantages:
Short Read Lengths (50-75 bp): The read lengths are relatively short, which can complicate the assembly of complex genomes and the analysis of repetitive regions.
High Startup Costs: The initial investment in Illumina sequencing equipment and infrastructure is substantial.
De Novo Assembly Can Be Challenging: Assembling genomes from short reads can be difficult, especially for organisms with complex or repetitive genomes.
Introduced by Mostafa Ronaghi and colleagues in 1996, 454 pyrosequencing involves the ligation of DNA fragments to adapters, followed by amplification through emulsion PCR (emPCR). The amplified fragments are then sequenced in picoliter-sized reaction chambers. DNA synthesis is monitored by detecting the release of pyrophosphate, which generates light in a reaction catalyzed by luciferase. The intensity of the light corresponds to the number of nucleotides incorporated, and this information is used to determine the sequence.
Advantages:
Low Error Rate: 454 pyrosequencing has a relatively low error rate compared to other sequencing methods.
Medium Read Length (400-700 bp): The read lengths are longer than those of many other NGS technologies, making it useful for certain applications.
Disadvantages:
High Cost per Base: The cost of sequencing with 454 technology is higher than that of some other methods.
Requires Large-Scale Operations: The technology is best suited for large-scale sequencing projects, which can limit its accessibility for smaller laboratories.
Ion Torrent technology, developed by Life Technologies, converts nucleotide sequences into digital information using a semiconductor chip. When a nucleotide is incorporated into a growing DNA strand, a hydrogen ion is released, causing a detectable change in pH. This change is recorded as a voltage shift, which is used to determine the sequence. The technology allows for real-time sequencing without the need for fluorescent labels.
Advantages:
Low Startup Costs: The initial investment in Ion Torrent technology is lower compared to some other NGS platforms.
Scalable Data Output: The technology can be scaled to produce varying amounts of data, making it flexible for different project sizes.
Fast Sequencing Runs: Ion Torrent sequencing is relatively fast, with runs taking only a few hours.
Disadvantages:
Short Read Lengths (100-200 bp): The read lengths are relatively short, which can limit its use in certain applications.
New Technology with Evolving Applications: As a newer technology, Ion Torrent sequencing is still evolving, and its full range of applications is still being explored.
Pacific Biosciences' single molecule real-time (SMRT) sequencing technology offers very long read lengths using a unique circular DNA molecule called a "SMRTbell." The DNA is synthesized in real-time, with each nucleotide labeled with a different fluorophore. The incorporation of each nucleotide is detected by imaging the fluorescent signal, which is timed with the rate of nucleotide incorporation.
Advantages:
Potential for Very Long Reads (Several kb+): PacBio sequencing can produce extremely long read lengths, which is beneficial for assembling complex genomes and analyzing repetitive regions.
Single-Molecule Sequencing: The technology sequences individual DNA molecules, providing a more accurate representation of the genome.
Disadvantages:
High Error Rate (10-15%): The error rate is relatively high, although it can be mitigated by high-coverage sequencing.
High Cost per Base: The cost of sequencing with PacBio technology is higher than some other methods.
High Startup Costs: The initial investment in PacBio sequencing equipment is substantial.
Oxford Nanopore Technologies' systems, such as MinION, use biological nanopores to identify nucleotide sequences by detecting changes in electrical conductivity as DNA strands pass through the pores. This technology is portable and can produce very long reads, making it suitable for fieldwork and real-time analysis.
Advantages:
Extremely Long Sequencing Reads: Nanopore sequencing can produce reads of several megabases, making it ideal for assembling large genomes and studying structural variations.
Short Processing Time: The technology allows for rapid sequencing, with minimal sample preparation time.
Portable and Suitable for Fieldwork: The small size of devices like MinION makes them highly portable, enabling sequencing in remote or resource-limited settings.
Disadvantages:
Limited Sequencing Accuracy: The accuracy of nanopore sequencing is lower than some other methods, although improvements are being made.
Context-Dependent Sequencing Bias: The accuracy and efficiency of sequencing can be affected by the context of the nucleotide sequence.
Not Yet Widely Adopted: As a relatively new technology, nanopore sequencing is not yet as widely adopted as other sequencing methods, and its protocols and applications are still being refined.
DNA sequencing has fundamentally transformed numerous scientific and clinical disciplines by providing detailed insights into the genetic blueprint of organisms. This technology has a wide range of applications across various fields, each benefiting uniquely from the ability to decode and understand genetic information.
Molecular Biology
In molecular biology, DNA sequencing is a critical tool for understanding the intricate details of genetic material. Researchers use sequencing to identify changes in genes, which can reveal mutations linked to diseases and phenotypes. This ability is pivotal for studying genetic disorders, cancer, and many other health conditions. Sequencing enables scientists to pinpoint specific genetic variations that may cause or contribute to diseases, facilitating the development of targeted therapies and personalized medicine. For example, by identifying mutations in cancer genes, researchers can develop drugs that specifically target these alterations, improving treatment efficacy and reducing side effects.
Moreover, DNA sequencing has revolutionized the study of gene expression and regulation. By analyzing the sequences of regulatory regions, scientists can understand how genes are turned on or off in different tissues and under various conditions. This knowledge is crucial for understanding developmental processes, cellular differentiation, and the mechanisms underlying many diseases. Additionally, sequencing allows for the identification of new genes and genetic elements, expanding our understanding of the genome's functional landscape.
Evolutionary Biology
DNA sequencing has had a profound impact on evolutionary biology by enabling the reconstruction of phylogenetic trees, which depict the evolutionary relationships between different organisms. By comparing the genetic sequences of various species, researchers can infer the evolutionary history and divergence times of these species. This information provides insights into how life has evolved over millions of years, shedding light on the processes of natural selection, adaptation, and speciation.
Sequencing also allows for the study of ancient DNA, providing a window into the genomes of extinct species. This has led to remarkable discoveries, such as the sequencing of Neanderthal and Denisovan genomes, which have revealed interbreeding events between these ancient hominins and modern humans. These insights have deepened our understanding of human evolution and the genetic contributions of these archaic humans to present-day populations.
Metagenomics
Metagenomics is the study of genetic material recovered directly from environmental samples, bypassing the need for culturing. This approach has opened new avenues for exploring microbial diversity and function in various environments, such as soil, water, and the human gut. By sequencing the collective genomes of microbial communities, researchers can identify the species present and their potential metabolic capabilities.
This information is vital for understanding ecosystem dynamics, nutrient cycling, and the roles of microbes in health and disease. For example, metagenomic studies of the human gut microbiome have revealed the vast diversity of microbial species living in our intestines and their contributions to digestion, immunity, and overall health. Changes in the gut microbiome composition have been linked to numerous conditions, including obesity, diabetes, and inflammatory bowel diseases, highlighting the importance of metagenomics in medical research.
Virology
In virology, DNA sequencing is an indispensable tool for identifying and studying viruses. Given the small size of viral genomes, sequencing is particularly suited for characterizing viral genetic material. This capability is crucial for understanding viral evolution, transmission, and pathogenicity. Sequencing can be used to track mutations in viral genomes, which can impact virulence, transmissibility, and resistance to antiviral drugs.
During viral outbreaks, sequencing provides valuable information about the origin and spread of the virus. By comparing viral sequences from different patients, researchers can trace the transmission pathways and identify potential sources of infection. This approach was instrumental during the COVID-19 pandemic, where sequencing efforts helped monitor the emergence of new variants and inform public health responses.
Medicine
The application of DNA sequencing in medicine has revolutionized diagnostics, treatment, and personalized healthcare. Sequencing is increasingly used to diagnose genetic disorders by identifying mutations responsible for conditions such as cystic fibrosis, muscular dystrophy, and various hereditary cancers. This enables early diagnosis and intervention, which can significantly improve patient outcomes.
In oncology, sequencing tumor DNA allows for the identification of specific genetic mutations driving cancer progression. This information can guide the selection of targeted therapies that are more effective and have fewer side effects compared to traditional treatments. Personalized medicine, which tailors treatment to an individual's genetic makeup, is becoming more feasible with advances in sequencing technology.
Sequencing is also crucial for infectious disease management. It can identify the specific pathogens causing an infection, allowing for more accurate and timely treatment with appropriate antibiotics or antivirals. This approach is essential for combating antibiotic resistance, as it ensures that patients receive the most effective treatment based on the genetic profile of the infecting organism.
Forensic Science
In forensic science, DNA sequencing is a powerful tool for identification and paternity testing. Sequencing can analyze DNA from various sources, such as fingerprints, saliva, hair follicles, and other biological samples. The unique genetic fingerprints provided by sequencing enable precise identification of individuals, which is invaluable in criminal investigations, disaster victim identification, and paternity disputes.
Sequencing can also help solve cold cases by analyzing previously untested or degraded DNA samples. Advances in sequencing technology have increased the sensitivity and accuracy of DNA analysis, making it possible to obtain genetic information from very small or damaged samples.
Whole Genome Sequencing (WGS) involves sequencing an organism's entire genome, providing a comprehensive view of its genetic makeup. WGS has broad applications across various fields:
Identifying Inherited Disorders in Newborns: WGS can detect genetic mutations responsible for inherited disorders, allowing for early diagnosis and intervention. This can significantly improve the quality of life for affected individuals and inform family planning decisions.
Tracking Disease Outbreaks: WGS enables the monitoring of disease outbreaks by identifying and tracking genetic changes in pathogens. This information can help public health officials implement effective containment strategies and develop vaccines.
Tailoring Treatments to Individual Genetic Profiles: WGS allows for personalized medicine by identifying genetic variations that affect drug response. This enables healthcare providers to select the most effective treatments with the fewest side effects for each patient.
Studying Mutations Driving Cancer Progression: WGS can identify mutations that contribute to cancer development and progression. This information can guide the development of targeted therapies and improve cancer treatment outcomes.
Characterizing Rare Cancer Genomes: WGS can provide insights into the genetic basis of rare cancers, which may have unique mutations not found in more common cancers. This can lead to the development of novel treatments and improve understanding of cancer biology.
Detecting Disease-Contributing Variants in Families: WGS can identify genetic variants that contribute to disease within families, allowing for more accurate risk assessment and early intervention.
Conducting Large-Scale Longitudinal Studies of Complex Traits: WGS is essential for studying complex traits influenced by multiple genetic and environmental factors. Longitudinal studies using WGS can identify genetic variants associated with these traits and provide insights into their underlying mechanisms.
The applications of DNA sequencing are vast and varied, spanning multiple disciplines and offering profound insights into genetics, biology, and medicine. As sequencing technologies continue to advance, their impact will only grow, driving further discoveries and innovations that will enhance our understanding of the natural world and improve human health
DNA sequencing has revolutionized scientific research, providing unparalleled insights into genetics, evolution, and disease. From Fredrick Sanger's foundational chain termination method to advanced next-generation sequencing technologies, each innovation has broadened the scope of genomic research. These advancements have transformed molecular biology, evolutionary biology, medicine, and forensic science by enabling precise genetic analysis, the reconstruction of evolutionary histories, the diagnosis and treatment of genetic disorders, and accurate forensic identification. Whole genome sequencing exemplifies the broad applications of sequencing technologies, facilitating the tracking of disease outbreaks, the study of cancer progression, and large-scale genetic studies. As sequencing technologies continue to evolve, they promise to deliver even more significant breakthroughs, further enhancing our understanding of the genetic underpinnings of life and driving innovations across multiple scientific disciplines.
DNA sequencing determines the order of DNA building blocks [Internet]. What is Biotechnology. Available from: https://www.whatisbiotechnology.org/index.php/science/summary/sequencing/dna-sequencing-determines-the-order-of-dna-building-blocks#:~:text=DNA%20sequencing%20is%20a%20method,to%20become%20(the%20phenotype).
DNA sequencing [Internet]. Khan Academy. Available from: https://www.khanacademy.org/science/ap-biology/gene-expression-and-regulation/biotechnology/a/dna-sequencing
DNA Sequencing: Definition, Methods, and Applications [Internet]. CD Genomics. Available from: https://www.cd-genomics.com/blog/dna-sequencing-definition-methods-and-applications/
Scientists Say: DNA sequencing [Internet]. Science News Explores. Available from: https://www.snexplores.org/article/scientists-say-dna-sequencing
Collins FS, Fink L. The Human Genome Project. Alcohol Health Res World. 1995;19(3):190-195.
First Complete Sequence of Human Genome [Internet]. National Institutes of Health (NIH). Available from: https://www.nih.gov/news-events/nih-research-matters/first-complete-sequence-human-genome
The Human Genome Project [Internet]. National Human Genome Research Institute (NHGRI). Available from: https://www.genome.gov/human-genome-project
Satam H, Joshi K, Mangrolia U, Waghoo S, Zaidi G, Rawool S, et al. Next-Generation Sequencing Technology: Current Trends and Advancements. Biology (Basel). 2023 Jul 13;12(7):997.
Henson J, Tischler G, Ning Z. Next-generation sequencing and large genome assemblies. Pharmacogenomics. 2012 Jun;13(8):901-15.
Next Generation Sequencing [Internet]. Illumina. Available from: https://sapac.illumina.com/science/technology/next-generation-sequencing.html
Brittain HK, Scott R, Thomas E. The rise of the genome and personalised medicine. Clin Med (Lond). 2017 Dec;17(6):545-551.
Cembrowska-Lech D, Krzemińska A, Miller T, Nowakowska A, Adamski C, Radaczyńska M, et al. An Integrated Multi-Omics and Artificial Intelligence Framework for Advance Plant Phenotyping in Horticulture. Biology (Basel). 2023 Sep 30;12(10):1298.
Ashraf MF, Hou D, Hussain Q, Imran M, Pei J, Ali M, et al. Entailing the Next-Generation Sequencing and Metabolome for Sustainable Agriculture by Improving Plant Tolerance. Int J Mol Sci. 2022 Jan 7;23(2):651.
Heather JM, Chain B. The sequence of sequencers: The history of sequencing DNA. Genomics. 2016 Jan;107(1):1-8. doi: 10.1016/j.ygeno.2015.11.003. Epub 2015 Nov 10. PMID: 26554401; PMCID: PMC4727787.
A Journey Through the History of DNA Sequencing [Internet]. The DNA Universe. Available from: https://the-dna-universe.com/2020/11/02/a-journey-through-the-history-of-dna-sequencing/
Sanger Sequencing: Introduction, Principle, and Protocol [Internet]. CD Genomics. Available from: https://www.cd-genomics.com/blog/sanger-sequencing-introduction-principle-and-protocol/
Sanger sequencing [Internet]. Wikipedia. Available from: https://en.wikipedia.org/wiki/Sanger_sequencing
Sanger Sequencing [Internet]. Sigma-Aldrich. Available from: https://www.sigmaaldrich.com/IN/en/technical-documents/protocol/genomics/sequencing/sanger-sequencing
What are the advantages and disadvantages of the following DNA sequencing method: Sanger sequencing? [Internet]. Study.com. Available from: https://homework.study.com/explanation/what-are-the-advantages-and-disadvantages-of-the-following-dna-sequencing-method-sanger-sequencing.html
Cheng C, Fei Z, Xiao P. Methods to improve the accuracy of next-generation sequencing. Front Bioeng Biotechnol. 2023 Jan 20;11:982111.
Al-Shuhaib MBS, Hashim HO. Mastering DNA chromatogram analysis in Sanger sequencing for reliable clinical analysis. J Genet Eng Biotechnol. 2023 Nov 13;21(1):115.
Maxam–Gilbert sequencing [Internet]. Wikipedia. Available from: https://en.wikipedia.org/wiki/Maxam%E2%80%93Gilbert_sequencing#:~:text=Maxam%E2%80%93Gilbert%20sequencing%20is%20a,adjacent%20to%20the%20modified%20nucleotides.
Heather JM, Chain B. The sequence of sequencers: The history of sequencing DNA. Genomics. 2016 Jan;107(1):1-8.
Maxam–Gilbert sequencing [Internet]. ScienceDirect. Available from: https://www.sciencedirect.com/topics/neuroscience/maxam-gilbert-sequencing
DNA Sequencing: Maxam-Gilbert and Sanger Dideoxy Method [Internet]. Microbe Notes. Available from: https://microbenotes.com/dna-sequencing-maxam-gilbert-and-sanger-dideoxy-method/
What is Shotgun Sequencing? [Internet]. Your Genome. Available from: https://www.yourgenome.org/theme/what-is-shotgun-sequencing/#:~:text=Shotgun%20sequencing%20involves%20randomly%20breaking,is%20quicker%20and%20less%20expensive.
Shotgun sequencing [Internet]. Wikipedia. Available from: https://en.wikipedia.org/wiki/Shotgun_sequencing
Shotgun Sequencing [Internet]. News-Medical.net. Available from: https://www.news-medical.net/life-sciences/Shotgun-Sequencing.aspx
ABI Solid Sequencing [Internet]. Wikipedia. Available from: https://en.wikipedia.org/wiki/ABI_Solid_Sequencing
Sequencing Technology [Internet]. Illumina. Available from: https://sapac.illumina.com/science/technology/next-generation-sequencing/sequencing-technology.html
Gupta N, Verma VK. Next-Generation Sequencing and Its Application: Empowering in Public Health Beyond Reality. In: Microbial Technology for the Welfare of Society. 2019 Sep 13;17:313-41.
Principle and Workflow of Illumina Next-Generation Sequencing [Internet]. CD Genomics. Available from: https://www.cd-genomics.com/blog/principle-and-workflow-of-illumina-next-generation-sequencing/
Siqueira JF Jr, Fouad AF, Rôças IN. Pyrosequencing as a tool for better understanding of human microbiomes. J Oral Microbiol. 2012;4.
Fakruddin M, et al. Pyrosequencing-A Next Generation Sequencing Technology. World Applied Sciences Journal. 2013;24(12):1558-1571.
Slatko BE, Gardner AF, Ausubel FM. Overview of Next-Generation Sequencing Technologies. Curr Protoc Mol Biol. 2018 Apr;122(1).
Ion Torrent Next Generation Sequencing Technology [Internet]. Thermo Fisher. Available from: https://www.thermofisher.com/in/en/home/life-science/sequencing/next-generation-sequencing/ion-torrent-next-generation-sequencing-technology.html
Ion semiconductor sequencing [Internet]. Wikipedia. Available from: https://en.wikipedia.org/wiki/Ion_semiconductor_sequencing
Nakano K, Shiroma A, Shimoji M, Tamotsu H, Ashimine N, Ohki S, et al. Advantages of genome sequencing by long-read sequencer using SMRT technology in medical area. Hum Cell. 2017 Jul;30(3):149-161.
How HiFi Sequencing Works [Internet]. PacBio. Available from: https://www.pacb.com/technology/hifi-sequencing/how-it-works/
Logsdon GA, Vollger MR, Eichler EE. Long-read human genome sequencing and its applications. Nat Rev Genet. 2020 Oct;21(10):597-614.
Sequel System [Internet]. Biocompare. Available from: https://www.biocompare.com/23967-Next-Generation-Sequencers/11182080-Sequel-System/
Lu H, Giordano F, Ning Z. Oxford Nanopore MinION Sequencing and Genome Assembly. Genomics Proteomics Bioinformatics. 2016 Oct;14(5):265-279.
Nanopore sequencing [Internet]. ScienceDirect. Available from: https://www.sciencedirect.com/topics/neuroscience/nanopore-sequencing
Technology [Internet]. Oxford Nanopore Technologies. Available from: https://nanoporetech.com/platform/technology
Chen P, Sun Z, Wang J, Liu X, Bai Y, Chen J, et al. Portable nanopore-sequencing technology: Trends in development and applications. Front Microbiol. 2023;14:1043967.
MacKenzie M, Argyropoulos C. An Introduction to Nanopore Sequencing: Past, Present, and Future Considerations. Micromachines (Basel). 2023 Feb 16;14(2):459.
Koboldt DC, Steinberg KM, Larson DE, Wilson RK, Mardis ER. The next-generation sequencing revolution and its impact on genomics. Cell. 2013 Sep 26;155(1):27-38.
Next-Generation Sequencing [Internet]. Illumina. Available from: https://sapac.illumina.com/science/technology/next-generation-sequencing.html
Hood L, Rowen L. The Human Genome Project: big science transforms biology and medicine. Genome Med. 2013;5:79.
Li H, Yang Y, Hong W, et al. Applications of genome editing technology in the targeted therapy of human diseases: mechanisms, advances and prospects. Sig Transduct Target Ther. 2020;5:1.
DNA Sequencing: Definition, Methods, and Applications [Internet]. CD Genomics. Available from: https://www.cd-genomics.com/blog/dna-sequencing-definition-methods-and-applications/
Whole Genome Sequencing [Internet]. Illumina. Available from: https://sapac.illumina.com/techniques/sequencing/dna-sequencing/whole-genome-sequencing.html#:~:text=It%20is%20ideal%20for%20discovery,changes%2C%20and%20large%20structural%20variants.
Brlek P, Bulić L, Bračić M, Projić P, Škaro V, Shah N, et al. Implementing Whole Genome Sequencing (WGS) in Clinical Practice: Advantages, Challenges, and Future Perspectives. Cells. 2024 Mar 13;13(6):504.
Whole genome sequencing [Internet]. Wikipedia. Available from: https://en.wikipedia.org/wiki/Whole_genome_sequencing
Download PDF
Download PDF
Download PDF
Download PDF
