DNA Sequencing

Definition

DNA sequencing is a method used to determine the precise order of the four nucleotide bases – adenine, guanine, cytosine and thymine - that make up a strand of DNA. These bases provide the underlying genetic basis (the genotype) for telling a cell what to do, where to go and what kind of cell to become (the phenotype). Nucleotides are not the only determinants of phenotypes, but are essential to their formation. Each individual and organism has a specific nucleotide base sequence.

The two scientists in the photograph are reading the genetic code for a DNA sample on a highlighted light board. Such analysis is usually done by a computer. Credit: National Cancer Institute.

Importance

DNA sequencing played a pivotal role in mapping out the human genome, completed in 2003, and is an essential tool for many basic and applied research applications today. It has for example provided an important tool for determining the thousands of nucleotide variations associated with specific genetic diseases, like Huntington's, which may help to better understand these diseases and advance treatment.

DNA sequencing also underpins pharmacogenomics. This is a relatively new field which is leading the way to more personalised medicine. Pharmacogenomics looks at how a person's individual genome variations affect their response to a drug. Such data is being used to determine which drug gives the best outcome in particular patients. Over 140 drugs approved by the FDA now include pharmacogenomic information in their labelling. Such labelling is not only important in terms of matching patients to their most appropriate drug, but also for working out what their drug dose should be and their level of risk in terms of adverse events. Individual genetic profiling is already being used routinely to prescribe therapies for patients with HIV, breast cancer, lymphoblastic leukaemia and colon cancer and in the future will be used to tailor treatments for cardiovascular disease, cancer, asthma, Alzheimer's disease and depression. Drug developers are also using pharmacogenomic data to design drugs which can be targeted at subgroups of patients with specific genetic profiles.

Discovery

Although scientists established DNA had a double helix structure in 1953, it was to be many more years before they could analyse DNA fragments. In part this reflected the fact that small DNA molecules contain several thousands of nucleotides and it was difficult to obtain large quantities of homogeneous DNA. Scientists also lacked the means to degrade DNA which they needed for sequence analysis.

A new chapter opened up in the 1960s with the emergence of techniques to sequence ribonucleic acid (RNA)s. Ray Wu, a Chinese American biologist based at Cornell University, published one of the first methods for sequencing DNA in 1970. Using highly labelled deoxynucleotides (single units of DNA) and DNA polymerase he found a way to sequence the terminal region of a DNA molecule. Critically, Wu's approach broke the DNA sequence down into several different components for analysis, thereby circumventing the need for large quantities of homogeneous DNA. Subsequently, in 1971, Wu demonstrated his method could sequence the ends of DNA in lambda phage, and two years later that it had the capacity to determine the sequence of any DNA.

Over the course of the 1970s Wu's method was modified by Fred Sanger at the Laboratory of Molecular Biology in Cambridge, UK. In 1975 Sanger, together with Alan Coulson, published what became known as the 'Plus and Minus' technique. This enabled the sequencing of up to 80 nucleotides in one go. Three years later, in 1977, Sanger and his colleagues announced another technique called the 'Sanger method' or 'dideoxy sequencing'. This made it possible to sequence much longer stretches of DNA very rapidly. Their approach appeared alongside the reporting of another technique by Allan Maxam and Walter Gilbert at Harvard University.

While the Maxam-Gilbert method initially proved the most popular, it soon fell out of favour because it necessitated the use of hazardous chemicals and radioisotopes. Added to this, the method it was difficult to scale-up and could not be used in standard molecular biology kits because of its technical complexity. By contrast, the Sanger method gained popularity because it was easier to use and more reliable. It was also amenable to automation, paving the way to the first generation of automated DNA sequencers. The first automated DNA sequencer was devised in 1986 by Leroy Hood and colleagues at the California Institute of Technology together with a team including Lloyd Smith and Michael and Tim Hunkapiller. These machines used capillary electrophoresis rather than gel electrophoresis using slabs.

Several new DNA sequencing methods and machines have been developed since the 1990s. These were built following the introduction of microfluidic separation devices which improved sample injection and speeded up separation times. Such innovations improved both the efficiency and accuracy of sequencing, allowing for high-throughput sequencing, and radically lowered the cost. Between 2001 and 2011 the cost of sequencing a genome shrank from $100 million to $10,000.

Application

DNA sequencing provides the means to know how nucleotide bases are arranged in a piece of DNA. Several methods have been developed for this process. These have four key steps. In the first instance DNA is removed from the cell. This can be done either mechanically or chemically. The second phase involves breaking up the DNA and inserting its pieces into vectors, cells that indefinitely self-replicate, for cloning. In the third phase the DNA clones are placed with a dye-labelled primer (a short stretch of DNA that promotes replication) into a thermal cycler, a machine which automatically raises and lowers the temperature to catalyse replication. The final phase consists of electrophoresis, whereby the DNA segments are placed in a gel and subjected to an electrical current which moves them. Originally the gel was placed on a slab, but today it is inserted into a very thin glass tube known as a capillary. When subjected to an electrical current the smaller nucleotides in the DNA move faster than the larger ones. Electrophoresis thus helps sort out the DNA fragments by their size. The different nucleotide bases in the DNA fragments are identified by their dyes which are activated when they pass through a laser beam. All the information is fed into a computer and the DNA sequence displayed on a screen for analysis.

The method developed by Sanger was pivotal to the international Human Genome Project. Costing over US$3 billion and taking 13 years to complete, this project provided the first complete Human DNA sequence in 2003. Data from the project provided the first means to map out the genetic mutations that underlie specific genetic diseases. It also opened up a path to more personalised medicine, enabling scientists to examine the extent to which a patient's response to a drug is determined by their genetic profile. The genetic profile of a patient's tumour, for example, can now be used to work out what is the most effective treatment for an individual. It is also hoped that in the future that knowing the sequence of a person’s genome will help work out a person's predisposition to certain diseases, such as heart disease, cancer and type II diabetes, which could pave the way to better preventative care.

Data from the the Human Genome Project has also helped fuel the development of gene therapy, a type of treatment designed to replace defective genes in certain genetic disorders. In addition, it has provided a means to design drugs that can target specific genes that cause disease.

Beyond medicine, DNA sequencing is now used for genetic testing for paternity and other family relationships. It also helps identify crime suspects and victims involved in catastrophes. The technique is also vital to detecting bacteria and other organisms that may pollute air, water, soil and food. In addition the method is important to the study of the evolution of different population groups and their migratory patterns as well as determining pedigree for seed or livestock.

DNA Sequencing: timeline of key events

A Swiss physician and biochemist. Miescher, was the first person to isolate nucleic acids which would subsequently be found to carry the genetic blueprint for life. 1895-08-26T00:00:00+0000Parents Frederick and Cicely Sanger (nee Crewdson)1918-08-13T00:00:00+0000The first to determine the DNA sequence of insulin, Sanger proved proteins have a defined chemical composition. He was also pivotal to the development of the dideoxy chain-termination method for sequencing DNA molecules, known as the Sanger method. This provided a breakthrough in the sequencing of long stretches of DNA in terms of speed and accuracy and laid the foundation for the Human Genome Project. 1918-08-13T00:00:00+0000Werner Arber shared the 1978 Nobel Prize in Medicine for the discovery of 'restriction enzymes and their application to problems in molecular genetics.'1929-06-03T00:00:00+00001932-01-01T00:00:00+0000Studies a combination of chemistry, physics, maths and physiology and specialises in biochemistry in his final year.1936-01-01T00:00:00+0000Initially supervised by Bill Pirie, and then by Albert Neuberger, in the Department of Biochemistry. Thesis: 'On the metabolism of the amino acid lysine in the animal body'. 1940-01-01T00:00:00+0000Sanger undertakes the research as part of team working with Albert Chibnall in Department of Biochemistry. His work is initially supported by a Beit Memorial Fellowship from 1944 and then by Medical Research Council from 1951. 1944-01-01T00:00:00+0000An American biochemist, Mullen was jointly awarded 1993 Nobel Prize in Chemistry for development of polymerase chain reaction (PCR) method to quickly make a huge number of copies of specific pieces of DNA.1944-12-28T00:00:00+0000Venter founded Celera Genomics and The Institute for Genomic Research and J Craig Institute which helped sequence the first human genome. It was mostly his genome that was sequenced. 1946-10-14T00:00:00+0000Noted by Salvador Luria and his graduate student Mary Human while conducting experiments into the break-up of DNA in phage-infected bateria.1952-01-01T00:00:00+0000Sanger's insulin results establish for the first time that proteins are chemical entities with a defined sequence. The technique Sanger develops for sequencing insulin later becomes known as the degradation or DNP method. It provides the basis for his later development of sequencing tecdhniques for nucleic acids, including RNA and DNA.1955-01-01T00:00:00+0000Ingram shows that the difference between sickle-cell and normal haemoglobulin lies in just one amino acid. 1957-01-01T00:00:00+0000Prize awarded to Sanger 'for his work on the structure of proteins, especially that of insulin'.1958-01-01T00:00:00+0000Non-profit institution founded by Robert S Ledley to explore the use of computers in biomedical research. It is eventually located at Georgetown University Medical Center in Washington, D.C.1960-01-01T00:00:00+00001960-01-01T00:00:00+0000Werner Arber, Swiss microbiologist and geneticist, and his doctoral student Daisy Dussoix propose bacteria produce restriction and modification enzymes to counter invading viruses. 1962-01-01T00:00:00+0000Sanger now has close contact with protein crystallographers, molecular geneticists and protein chemists1962-01-01T00:00:00+0000Robert Holley and colleagues sequence Escherichia coli alanine transfer RNA, laying the foundation for DNA sequencing. 1965-01-01T00:00:00+0000W. Arber, 'Host-controlled modification of bacteriophage', Annual Review Microbiology 19 (1965), 365-78.1965-01-01T00:00:00+0000Book contains all protein sequences known to-date. It is the result of a collective effort to co-ordinate the ever-growing amount of information about protein sequences and their biochemical function. 1965-01-01T00:00:00+0000900 page monograph provides the first introduction to the application of digital computing in biology and medicine. 1965-01-01T00:00:00+0000Tested on ribosomal RNA1965-01-01T00:00:00+0000The sequencer was developed by Pehr Victor Edman with Geoffrey Begg1967-01-01T00:00:00+0000Discovery lays foundation for recombinant DNA and DNA sequencing.1968-01-01T00:00:00+0000Ray Wu and A.D. Kaiser report on the partial sequence of bacteriophage lambda DNA in the Journal of Molecular Biology, 35/3 (1968), 523-37. 1968-01-01T00:00:00+0000Kjell Kleppe, a Norwegian scientist working in H. Gobind Khorana's Institute for Enzyme Research at University of Wisconsin publishes papers describing the principles of PCR.1969-01-01T00:00:00+0000Called Thermus aquaticus (Taq) this enzyme becomes a standard source of enzymes because it can withstand higher temperatures than those from E Coli. Taq is later important in the PCR technique. 1969-01-01T00:00:00+0000Hamilton O Smith, Kent W Wilcox, Journal of Molecular Biology 51/2 (1970), 379-91. 1970-07-01T00:00:00+0000K. Kleppe, E Ohtsuka, R Kleppe, I Molineux, HG Khorana, "Studies on polynucleotides *1, *2XCVI. Repair replication of short synthetic DNA's as catalyzed by DNA polymerases", Journal of Molecular Biology, 56/2 (1971), 341-61. The method provides an artificial system of primers and templates that allows DNA polymerase to copy segments of the gene being synthesised. 1971-01-01T00:00:00+0000The 12 base sequence of bacteriophage lambda DNA is published by Ray Wu and Ellen Taylor in the Journal of Molecular Biology, 57 (1971) 0, 491-511. 1971-05-01T00:00:00+0000Kathleen Danna and Daniel Nathans, PNAS, 68/12 (1971), 2913-17.1971-12-01T00:00:00+0000This is achieved by Walter Gilbert and Allan Maxam at Harvard University using a method known as wandering-spot analysis.1973-01-01T00:00:00+0000The method enables 80 nucleotides to be sequenced in one go. Represents radical new approach which allows direct visual scanning of a sequence. 1975-01-01T00:00:00+0000This is found to contain 5,385 nucleotides. It is the first DNA based organism to have its complete genome sequenced. Sanger and his team use the plus and minus technique to determine the sequence. 1977-01-01T00:00:00+0000Duncan McCallum, a business computer programmer in Cambridge writes the first computer programme for DNA sequencing. This is used by Sanger's sequencing group at the MRC Laboratory of Molecular Biology. 1977-01-01T00:00:00+0000Two separate teams, one led by Fred Sanger at the MRC Laboratory of Molecular Biology, Cambridge, UK, and one composed of Allan Maxam, and Walter Gilbert at Harvard University publish two different methods for sequencing DNA. The first, known as the Sanger Method, or dideoxy sequencing, involves the breaking down and then building up of DNA sequences. The second, the Maxam-Gilbert method, involves the partial chemical modification of nucleotides in DNA. 1977-02-01T00:00:00+0000Awarded to Werner Arber, Daniel Nathans and Hamilton O Smith.1978-10-01T00:00:00+0000Prize shared with Walter Gilbert. Awarded on the basis of their 'contributions concerning the determination of base sequences in nucleic acids.' 1980-01-01T00:00:00+0000The aim is to establish a centralised sequence computerised database tha is available free of charge. 1980-01-01T00:00:00+0000The database was developed by Margaret Dayhoff, an American physical chemist, for storing nucleic acid sequences. It was a sophisticated on-line computer database that was accessible by telephone to outside users.1980-09-01T00:00:00+0000Database was started by Margaret Dayhoff at the NBRF in the mid 1960s and comprised over 200,000 residues. Within a month of its operation more than 100 scientists had requested access to the database. The database was funded with contributions from m Genex, Merck, Eli Lilly, DuPont, Hoffman–La Roche, and Upjohn, and computer time donated by Pfizer Medical Systems.1980-09-01T00:00:00+0000In this method genomic DNA is randomly fragmented and cloned to produce a random library in E Coli. The clones are then sequenced at random and the results assembled by computer which compares all of the sequence reads and aligns the matching sequences to produce the complete genome sequence. 1982-01-01T00:00:00+0000Funding secured for the setting up of GenBank, to be located at Los Alamos National Laboratory. It was to serve as a repository for newly determined sequences, as a tool for sequencers assembling genomes and for bioinformatic researchers. 1982-06-01T00:00:00+00001983-01-01T00:00:00+0000Kary Mullis, an American biochemist based at Cetus, proposed an alternative method to Sanger's DNA sequencing method to analyse Sickle cell Anaemia mutation which laid the foundation for the development of the PCR technique. 1983-05-01T00:00:00+0000Mullis reports on his production of olgionucleotides and some results from his experiments with PCR to Cetus Corporation's annual meeting but few show any interest. 1984-06-01T00:00:00+0000The application establishes polymerase chain reaction (PCR) as a method for amplifying DNA in vitro. PCR uses heat and enzymes to make unlimited copies of genes and gene fragments. The application is broad and is based on analysis of Sickle Cell Anaemia mutation via PCR and Oligomer restriction. 1985-03-01T00:00:00+0000Alec Jeffreys published hypervariable minisatellite regions in human DNA, which paved the way to DNA fingerprinting.1985-03-07T00:00:00+0000The technique enabled the amplification of small fragments of DNA on a large scale. It was published in RK Saiki et al, Enzymatic Amplification of beta-globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell Anemia, Science, 230 (1985), 1350-54.1985-12-20T00:00:00+0000Leroy Hood and colleagues at the California Institute of Technology together with a team including Lloyd Smith and Michael and Tim Hunkapiller, develop the first automated DNA sequencing machine. The machine is commercialised by Applied Biosystems. 1986-01-01T00:00:00+0000 First meeting on genome mapping and sequencing takes place at Cold Spring Harbor.1986-04-30T00:00:00+0000Funding secured for precursor of the Human Genome Project. US$10.7 million provided by Department of Energery and US$17.2 million by National Institutes of Health.1988-01-01T00:00:00+0000This method, called FASTA, is published by William R Pearson and David J Lipman in Proc Natl Acad Sci USA, 85/8 (April 1988), 2444-8. This is now a common tool for bioinformatics. It allos for the comparison and aligning of sequences. 1988-04-01T00:00:00+0000Joint working group of the US Department of Energy and the National Institututes of Health present plan Understanding Our Genetic Inheritance: The US Human Genome Project.1990-02-01T00:00:00+0000International scientific collaboration, initiated by the US Department of Energy, begun to determine the sequence of chemical base pairs which make up DNA, and to identify and map approximately 20,000 to 25,000 genes of the human genome. 1990-10-01T00:00:00+00001992-01-01T00:00:00+0000M. Frommer, L.E. McDonald, D.S. Millar, C.M. Collis, F. Watt, G.W. Grigg, P.L. Molloy, C.L. Paul, 'A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands', PNAS, 89/5 (1992), 1827-31.1992-03-01T00:00:00+00001995-07-28T00:00:00+00001996-01-01T00:00:00+0000Mostafa Ronaghi and Pal Nyren at the Royal Institute of Technology in Stockholm develop pyrosequencing which allows for shotgun sequencing without cloning in E coli or any host cell. The marchinery and reagents involved in the method was first commercialised by Pyrosequencing AB.1996-01-01T00:00:00+0000Celera Corporation launches a parallel effort to sequence the human genome to the Human Genome Project. Celera's entry into the field pose policy concerns about open access to gene sequencing data and accelerates the sequencing process in the Human Genome Project. 1998-05-01T00:00:00+0000The full 97 Mbp genome of Caenorhabditis elegans is found to have more than 19,000 genes.1998-12-11T00:00:00+0000Sequence of the first human chromosome (22) is published. 1999-01-01T00:00:00+00002000-01-01T00:00:00+0000U.S. President Bill Clinton and the British Prime Minister Tony Blair announced the completion of a rough draft of the human genome. 2000-06-26T00:00:00+0000Map published for the genome of the flowering plant Arabidopsis thaliana. The sequenced genome contains 25,498 genes encoding proteins from 11,000 families.2000-12-14T00:00:00+0000A consortium including scientists from Celera Genomics and 13 other organizations publishes the first consensus sequence of human genome. It is shown to have a 2.91 billioon base pair sequence. 2001-02-16T00:00:00+00002002-01-01T00:00:00+0000Researchers at the State University of New York report the creation of a full-genome complementary DNA from published sequences of poliovirus, and its reverse transcription into viral RNA2002-07-11T00:00:00+0000Genomic sequence completed for Plasmodium falciparum, the malaria parasite, which carries some 5,300 genes (Celera Genomics) and for malaria Anopheles gambiae, mosquito's principal vector (TIGR and Sanger Centre). 2002-10-03T00:00:00+0000The Human Genome Project was completed, two years ahead of schedule and at a cost of US$2.7 billion. Most of the government-sponsored sequencing was performed in universities and research centres from the United States, the United Kingdom, Japan, France, Germany. 2003-04-14T00:00:00+0000US FDA approved Roche AmpliChip Cytochrome P450 Genotyping Test. The first DNA microarray system approved for clinical applications, AmpliChip is designed to match patients with their most effective drug and determine appropriate drug doses. 2004-12-23T00:00:00+0000 Germ-line cell experiments remain off-limit. Sequence of the last chromosome in the Human Genome Project is published in Nature.2006-05-01T00:00:00+00002011-01-01T00:00:00+0000Sharon Peacock and Julian Parkhill together with other researchers from the University of Cambridge and the Wellcome Trust Sanger Institute used whole genome sequencing to trace the spread of an outbreak of meticillin resistant Staphylococcus aureus (MRSA) in Rosie Hospital's special care baby unit. Prospective sequencing then led them to screen staff and identify the potential source of infection. The researchers reported that the cost of DNA sequencing for the infection was half of the 10,000 pounds spent by the hospital to combat the outbreak of MRSA.2012-06-01T00:00:00+0000Undertaken at the University of California's Rady Children's Hospital in San Diego, the study involves the sequencing of all the genes of individuals in 118 families with a neurodevelopment problem. 2012-12-01T00:00:00+00002013-11-19T00:00:00+0000Test developed to detect tumour DNA. Study investigated blood samples from 161 patients with stage 2 and 3 melanoma who had received surgery. Results showed that skin cancer was much more likely to return within a year of surgery in patients with faults in either BRAF or NRAS genes. R J Lee et al, 'Circulating tumor DNA predicts survival in patients with resected high-risk stage II/III melanoma', Annals of Oncology, mdx717, https://doi.org/10.1093/annonc/mdx7172017-11-03T00:00:00+0000Discovery made as a result of study of 177 members of the Old Order of Amish community in Indiana. S. Khan, et al, 'A null mutation in SERPINE1 protects against biological aging in humans', Science Advances, 3/11 (2017), DOI: 10.1126/sciadv.aao16172017-11-15T00:00:00+0000
Date Event People Places
26 Aug 1895Johann Friedrich Miescher diedMiescher 
13 Aug 1918Frederick Sanger, twice Nobel Prize winner, born in Rendcombe, United KingdomSangerRendcomb, Gloucestershire
13 Aug 1918Frederick Sanger, twice Nobel Prize winner, bornSangerRendcomb, Gloucestershire, United Kingdom
3 Jun 1929Werner Arber was born in Granichen, SwitzerlandArberUniversity of Geneva
1932Sanger attends Bryanston School, Dorset, as boarderSanger 
1936 - 1940Sanger takes degree in Natural Sciences at Cambridge UniversitySangerCambridge University
1940 - 1943Sanger studies for a doctorate at Cambridge UniversitySangerCambridge University
1944Sanger starts working on amino acid composition of insulinSangerCambridge University
December 1944Kary Banks Mullis was bornCetus Corporation
14 Oct 1946J Craig Venter was born in Salt Lake City, UtahSalt Lake City, Utah
1952First observation of the modification of viruses by bacteriaLuria, HumanUniversity of Illinois
1955Sanger completes the full sequence of amino acids in insulinSangerCambridge University
1957Victor Ingram breaks the genetic code behind sickle-cell anaemia using Sanger's sequencing techniqueIngram, SangerCambridge University
1958Sanger awarded his first Nobel Prize in ChemistrySangerCambridge University
1960National Biomedical Research Foundation establishedLedleyGeorgetown University
1960Sanger begins to devise ways to sequence nucleic acids, starting with RNASangerCambridge University
1962Concept of restriction and modification enzymes bornArber, DussoixUniversity of Geneva
1962Sanger moves to the newly created Laboratory of Molecular Biology in CambridgeSangerLaboratory of Molecular Biololgy
1965Transfer RNA is the first nucleic acid molecule to be sequencedHolleyCornell University
1965Werner Arber predicts restriction enzymes could be used as a labortory tool to cleave DNAArberUniversity of Geneva
1965Atlas of Protein Sequence and Structure publishedDayhoffNational Biomedical Research Foundation
1965Ledley publishes Uses of Computers in Biology and MedicineLedleyNational Biomedical Research Foundation
1965Sanger and colleagues publish two-dimension partition sequencing methodSanger, Brownlee, BarrellLaboratory of Molecular Biology
1967First automatic protein sequencer developedEdman, BeggSt Vincent's School of Medical Research
1968 - 1970Restriction enzymes found to act as chemical knives to cut DNASmith, NathansUniversity of Geneva, University of California in Berkeley, Johns Hopkins University
1968The first partial sequence of a viral DNA is reportedWu, KaiserCornell University, Stanford University Medical School
1969First principles for PCR publishedKhorana, KleppeUniversity of Wisconsin-Madison
1969New species of bacterium is isolated from hot spring in Yellowstone National Park by Thomas BrockBrockCase Western Reserve University
July 1970First restriction enzyme isolated and characterisedSmith, WilcoxJohns Hopkins University
1971Process called repair replication for synthesising short DNA duplexes and single-stranded DNA by polymerases is publishedKhorana, KleppeMIT
May 1971Complete sequence of bacteriophage lambda DNA reportedWu, TaylorCornell University
December 1971First experiments published demonstrating the use of restriction enzymes to cut DNADanna, NathansJohns Hopkins University
1973The sequencing of 24 basepairs is reportedGilbert, MaxamHarvard University
1975Sanger and Coulson publish their plus minus method for DNA sequencingSanger, CoulsonLaboratory of Molecular Biology
1977Complete sequence of bacteriophage phi X174 DNA determinedSangerLaboratory of Molecular Biology
1977First computer programme written to help with the compilation and analysis of DNA sequence dataMcCallumLaboratory of Molecular Biology
February 1977Two different DNA sequencing methods published that allow for the rapid sequencing of long stretches of DNASanger, Maxam, GilbertHarvard University, Laboratory of Molecular Biology
October 1978Nobel Prize for discovery and understanding of restriction enzymesArber, Nathans, SmithJohns Hopkins University, University of Geneva
1980Sanger awarded his second Nobel Prize in ChemistrySanger, GilbertHarvard University, Laboratory of Molecular Biology
January 1980European Molecular Biology Laboratory convenes meeting on Computing and DNA SequencesEMBL
September 1980First DNA sequence database createdDayhoffNational Biomedical Research Foundation
1980Largest nucleic acid sequence database in the world made available free over telephone networkDayhoffNational Biomedical Research Foundation
1982Whole genome sequencing method is introduced for DNA sequencing 
June 1982NIH agrees to provide US$3.2 million over 5 years to establish and maintain a nucleic sequence database 
1983Sanger retiresSangerLaboratory of Molecular Biology
1983Polymerase chain reaction (PCR) starts to be developed as a technique to amplify DNAMullisCetus Corporation
June 1984Results from PCR experiments start being reportedMullisCetus Corporation
March 1985Mullis and Cetus Corporation filed patent for the PCR techniqueMullisCetus Corporation
March 1985DNA fingerprinting principle laid outJeffreysUniversity of Leicester
20 Dec 1985The Polymerase Chain Reaction technique was publishedMullisCetus Corporation
1986First machine developed for automating DNA sequencingHood, Smith, HunkapillerCalifornia Institute of Technology, Applied Biosystems
30 Apr 1986Human Genome Organization founded 
1988US Congress funds genome sequencing 
April 1988Development of first rapid search computer programme to identify genes in a new sequencePearson, Lipman 
1 Feb 1990First pitch for US Human Genome Project 
October 1990Human Genome Project formally launched 
1992GenBank is integrated into the NIH National Center for Biotechnology Information 
1 Mar 1992Method devised to isolate methylated cytosine residues in individual DNA strands providing avenue to undertake DNA methylation genomic sequencing 
July 1995Craig Venter's team at The Institute of Genomics Research (TIGR) published the first complete sequence of the 1.8 Mbp genome of a free-living organism (the bacterium Haemophilus influenzae)VenterThe Institute for Genomic Research
1996Complete genome sequence of the first eukaryotic organism, the yeast S. cerevisiae, is published  
1996Pyrosequencing is introduced for DNA sequencingRonaghi, NyrenRoyal Institute of Technology
May 1998Commercial Human Genome Project launchedVenterCelera Genomics
December 1998Complete genome sequence of the first multicellular organism, the nematode worm Caenorhabditis elegans, is publishedSanger Institute, Washington University
1999First human chromosome sequence published 
2000Complete sequences of the genomes of the fruit fly Drosophila and the first plant, Arabidopsis, are published 
June 2000Human genome draft sequence announced 
December 2000First plant DNA sequenced 
February 2001First consensus sequence of human genome publishedCelera
2002Complete genome sequence of the first mammalian model organism, the mouse, is published  
July 2002Poliovirus synthesisedStony Brook University
October 2002Genomic sequence of the principal malaria parasite and vector reported 
April 2003The sequence of the first human genome was published 
23 Dec 2004First DNA microarray diagnostic device approved 
May 2006Last human chromosome is sequenced 
January 2011DNA sequencing proves useful to documenting the rapid evolution of Streptococcus penumoniae in response to the application of vaccinesWellcome Trust Sanger Institute
June 2012DNA sequencing helps identify the source of an MRSA outbreak in a neornatal intensive care unitPeacock, ParkhillCambridge University, Wellcome Trust Sanger Institute
December 2012DNA sequencing utilised for identifying neurological disease conditions different from those given in the original diagnosisUniversity of California San Diego
19 Nov 2013Sanger, the inventor of DNA sequencing, died at the age of 95SangerCambridge
3 Nov 2017Research published demonstrating simple blood test can identify which patients are at most risk of skin cancer returningLee, Gremel, Marshall, Myers, Fisher, Dunn, Dhomen, Corrie, Middleton, Lorigan, MaraisUniversity of Manchester
15 Nov 2017Rare mutation of gene called Serpine 1 discovered to protect against biological ageing processKhan, Shah, Klyachko, Baldridge, Eren, Place, Aviv, Puterman, Lloyd-Jones, Heiman, Miyata, Gupta, Shapiro, VaughanNorthwestern University, University of British Columbia, New Jersey Medical School, Tohoku University,

26 Aug 1895

Johann Friedrich Miescher died

13 Aug 1918

Frederick Sanger, twice Nobel Prize winner, born in Rendcombe, United Kingdom

13 Aug 1918

Frederick Sanger, twice Nobel Prize winner, born

3 Jun 1929

Werner Arber was born in Granichen, Switzerland

1932

Sanger attends Bryanston School, Dorset, as boarder

1936 - 1940

Sanger takes degree in Natural Sciences at Cambridge University

1940 - 1943

Sanger studies for a doctorate at Cambridge University

1944

Sanger starts working on amino acid composition of insulin

Dec 1944

Kary Banks Mullis was born

14 Oct 1946

J Craig Venter was born in Salt Lake City, Utah

1952

First observation of the modification of viruses by bacteria

1955

Sanger completes the full sequence of amino acids in insulin

1957

Victor Ingram breaks the genetic code behind sickle-cell anaemia using Sanger's sequencing technique

1958

Sanger awarded his first Nobel Prize in Chemistry

1960

National Biomedical Research Foundation established

1960

Sanger begins to devise ways to sequence nucleic acids, starting with RNA

1962

Concept of restriction and modification enzymes born

1962

Sanger moves to the newly created Laboratory of Molecular Biology in Cambridge

1965

Transfer RNA is the first nucleic acid molecule to be sequenced

1965

Werner Arber predicts restriction enzymes could be used as a labortory tool to cleave DNA

1965

Atlas of Protein Sequence and Structure published

1965

Ledley publishes Uses of Computers in Biology and Medicine

1965

Sanger and colleagues publish two-dimension partition sequencing method

1967

First automatic protein sequencer developed

1968 - 1970

Restriction enzymes found to act as chemical knives to cut DNA

1968

The first partial sequence of a viral DNA is reported

1969

First principles for PCR published

1969

New species of bacterium is isolated from hot spring in Yellowstone National Park by Thomas Brock

Jul 1970

First restriction enzyme isolated and characterised

1971

Process called repair replication for synthesising short DNA duplexes and single-stranded DNA by polymerases is published

May 1971

Complete sequence of bacteriophage lambda DNA reported

Dec 1971

First experiments published demonstrating the use of restriction enzymes to cut DNA

1973

The sequencing of 24 basepairs is reported

1975

Sanger and Coulson publish their plus minus method for DNA sequencing

1977

Complete sequence of bacteriophage phi X174 DNA determined

1977

First computer programme written to help with the compilation and analysis of DNA sequence data

Feb 1977

Two different DNA sequencing methods published that allow for the rapid sequencing of long stretches of DNA

Oct 1978

Nobel Prize for discovery and understanding of restriction enzymes

1980

Sanger awarded his second Nobel Prize in Chemistry

Jan 1980

European Molecular Biology Laboratory convenes meeting on Computing and DNA Sequences

Sep 1980

First DNA sequence database created

1980

Largest nucleic acid sequence database in the world made available free over telephone network

1982

Whole genome sequencing method is introduced for DNA sequencing

Jun 1982

NIH agrees to provide US$3.2 million over 5 years to establish and maintain a nucleic sequence database

1983

Sanger retires

1983

Polymerase chain reaction (PCR) starts to be developed as a technique to amplify DNA

Jun 1984

Results from PCR experiments start being reported

Mar 1985

Mullis and Cetus Corporation filed patent for the PCR technique

Mar 1985

DNA fingerprinting principle laid out

20 Dec 1985

The Polymerase Chain Reaction technique was published

1986

First machine developed for automating DNA sequencing

1986

Human Genome Organization founded

1988

US Congress funds genome sequencing

Apr 1988

Development of first rapid search computer programme to identify genes in a new sequence

Apr 1988

First pitch for US Human Genome Project

Oct 1990

Human Genome Project formally launched

1992

GenBank is integrated into the NIH National Center for Biotechnology Information

1 Mar 1992

Method devised to isolate methylated cytosine residues in individual DNA strands providing avenue to undertake DNA methylation genomic sequencing

Jul 1995

Craig Venter's team at The Institute of Genomics Research (TIGR) published the first complete sequence of the 1.8 Mbp genome of a free-living organism (the bacterium Haemophilus influenzae)

1996

Complete genome sequence of the first eukaryotic organism, the yeast S. cerevisiae, is published

1996

Pyrosequencing is introduced for DNA sequencing

May 1998

Commercial Human Genome Project launched

Dec 1998

Complete genome sequence of the first multicellular organism, the nematode worm Caenorhabditis elegans, is published

1999

First human chromosome sequence published

2000

Complete sequences of the genomes of the fruit fly Drosophila and the first plant, Arabidopsis, are published

Jun 2000

Human genome draft sequence announced

Dec 2000

First plant DNA sequenced

Feb 2001

First consensus sequence of human genome published

2002

Complete genome sequence of the first mammalian model organism, the mouse, is published

Jul 2002

Poliovirus synthesised

Oct 2002

Genomic sequence of the principal malaria parasite and vector reported

Apr 2003

The sequence of the first human genome was published

Apr 2003

First DNA microarray diagnostic device approved

May 2006

Last human chromosome is sequenced

Jan 2011

DNA sequencing proves useful to documenting the rapid evolution of Streptococcus penumoniae in response to the application of vaccines

Jun 2012

DNA sequencing helps identify the source of an MRSA outbreak in a neornatal intensive care unit

Dec 2012

DNA sequencing utilised for identifying neurological disease conditions different from those given in the original diagnosis

19 Nov 2013

Sanger, the inventor of DNA sequencing, died at the age of 95

3 Nov 2017

Research published demonstrating simple blood test can identify which patients are at most risk of skin cancer returning

15 Nov 2017

Rare mutation of gene called Serpine 1 discovered to protect against biological ageing process