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Sequencing Glossary & taxonomy
Evolving Terminologies for Emerging Technologies

Comments? Suggestions Revisions?
Mary Chitty MSLS 
mchitty@healthtech.com
Last revised January 07, 2020.



The "race" to sequence the Human Genome was not a 100 yard dash, but a marathon.  Although the Human Genome Project finished well ahead of schedule, and a number of genes have been identified, we have just begun to get a glimpse of what specific genes do and how we might be able to better use this knowledge for therapeutic interventions.  Teasing apart the interactions of  genes and proteins, delineating changes throughout the cell cycle, and correlating changes with health and disease will take even more time.  But with complete sequences, and the cross- species comparisons we can expect new insights and speeding up over time. Sequencing DNA is only a first step towards finding what functions are connected with specific sequences. Sequencing proteins (and determining the structures  – and functions of proteins) is ongoing.  

Related glossaries include  Biomarkers   Molecular Diagnostics   Molecular Medicine 
Informatics Bioinformatics  Drug discovery informatics  Sequencing informatics terms  in Genomic informatics
Technologies Chromatography & electrophoresis    Microarrays     Genomic technologies 
Biology 
Functional genomics   Genomics  Pharmacogenomics 
Proteins   Protein Structures    Proteomics  SNPs & genetic variations     Sequences - DNA & beyond    

$1,00 genome : Molecular Diagnostics

Clinical genome sequencing explores the recent surge in clinical genome sequencing, from the point of view of the sequencing providers, the medical organizations delivering these services, and the start-ups offering a variety of interpretation services, platforms, and business models. Aspects include: Progress in clinical genome sequencing, Organizations leading the way in generating clinical data and its interpretation, Determining the causality of documented variants in genetic disease, Clinical genome sequencing in oncology, Academic and commercial clinical genomics providers, next-gen sequencing landscape, Companies providing genome interpretation software, Initiatives in setting sequencing standards, custom survey results on clinical genome sequencing. Insight Pharma Reports Advances in Clinical Genome Sequencing and Diagnostics 2013   

Clinical NGS Diagnostics 2019 March 11-13 San Francisco CA https://www.triconference.com/Clinical-NGS-Diagnostics Clinical sequencing is enabling personalized medicine to combat a host of diseases, from cancers to infections. This program will discuss applications of circulating tumor cells, liquid biopsy utilization, and the technologies and approaches to strategize and optimize processes to bring developments to the clinic and beyond.

coverage [sequencing]:
Coverage is the average number of reads representing a given nucleotide in the reconstructed sequence ... Sometimes a distinction is made between sequence coverage and physical coverage. Sequence coverage is the average number of times a base is read (as described above). Physical coverage is the average number of times a base is read or spanned by mate paired reads[8] Wikipedia shotgun sequencing accessed Jan 10, 2011 http://en.wikipedia.org/wiki/Shotgun_sequencing#Coverage 

de novo sequencing: Determination of sequences (of genes or amino acids) whose sequence is not yet known. Can be done with LC/MS/MS or nanoelectrospray MS/MS.

From the Latin "de novo" from the beginning. See also Mass spectrometry

deep sequencing: Techniques of nucleotide sequence analysis that increase the range, complexity, sensitivity, and accuracy of results by greatly increasing the scale of operations and thus the number of nucleotides, and the number of copies of each nucleotide sequenced. The sequencing may be done by analysis of the synthesis or ligation products, hybridization to preexisting sequences, etc. MeSH 2011    

refers to the general concept of aiming for high number of unique reads of each region of a sequence.[3]  Wikipedia accessed 2018 Aug 28  https://en.wikipedia.org/wiki/Coverage_(genetics)

Deep Sequencing and Single Cell Analysis for Antibody Discovery Technologies and Best Practices for Applying Repertoire Analysis in the Discovery of Therapeutic Proteins JANUARY 23-24, 2020 San Diego CA  The rapid adoption of deep sequencing and single B cell analysis has given discovery scientists an extraordinary view into human and animal immune repertoires that is now informing all aspects of biopharmaceutical R&D. This dynamic field is bringing together the disciplines of immunology, structural and computational biology, informatics and microfluidics to offer previously unimaginable perspectives that will drive discovery of the next generation of biologic drugs. https://www.chi-peptalk.com/antibody-discovery

exome sequencing: Targeted sequencing of all protein-coding regions in the human genome -- now offers an unprecedented opportunity for systematic, genome-wide discovery of somatic mutations in tumor tissue. Cancer. New epigenetic drivers of cancers, Elsässer SJ, Allis CD, Lewis PW. Science. 2011 Mar 4;331(6021):1145-6. 
Wikipedia http://en.wikipedia.org/wiki/Exome_sequencing

genotype: The genetic constitution of an organism as revealed by genetic or molecular analysis, i.e. the complete set of genes, both dominant and recessive, possessed by a particular cell or organism. IUPAC Biotech

The observed alleles at a genetic locus for an individual. NHLBI    The genetic constitution of the individual; the characterization of the genes.  MeSH 1968

genotyping:  The determination of relevant nucleotide- base sequences in each of the two parental chromosomes. May refer to identifying one or more, up to the entire gene sequence of an organism. Compare phenotype. Used for diagnosis, drug efficacy, and toxicity. Utilizes genomic DNA that, after digestion, reacts with a SNP array to obtain an individual SNP pattern. These variations can for instance provide information about the diagnosis of a certain disease, or the effectiveness or side effect of a certain drug.

Genotyping implies (though I haven't found this in print) determining known variants, as opposed to discovery of new ones. Related terms SNPS & other genetic variations; Broader term sequencing; Narrower terms: haplotyping, genome wide association studies

What is the difference between genotyping and sequencing? 23andme
https://customercare.23andme.com/hc/en-us/articles/202904600-What-is-the-difference-between-genotyping-and-sequencing

GWAS Genome Wide Association Sequencing: Genomic informatics

haplogroups: The term 'haplogroup' refers to the SNP/unique-event polymorphism (UEP) mutations that represent the clade to which a collection of particular human haplotypes belong. (Clade here refers to a set of haplotypes sharing a common ancestor.)[7] A haplogroup is a group of similar haplotypes that share a common ancestor with a single-nucleotide polymorphism mutation.[8][9] Mitochondrial DNA passes along a maternal lineage that can date back thousands of years.[8]       Wikipedia accessed 2018 Nov 8    https://en.wikipedia.org/wiki/Haplotype

 (haploid from the Greek: ἁπλούς, haploûs, "onefold, simple" and English: group) is a group of similar haplotypes that share a common ancestor with a single-nucleotide polymorphism mutation.[3][4] More specifically, a haplogroup is a combination of alleles at different chromosomes regions that are closely linked and that tend to be inherited together. As a haplogroup consists of similar haplotypes, it is usually possible to predict a haplogroup from haplotypes. Haplogroups pertain to a single line of descent. As such, membership of a haplogroup, by any individual, relies on a relatively small proportion of the genetic material possessed by that individual.  Wikipedia accessed 2018 Nov 8 http://en.wikipedia.org/wiki/Haplogroup 

haplotype: The genetic constitution of individuals with respect to one member of a pair of allelic genes, or sets of genes that are closely linked and tend to be inherited together such as those of the MAJOR HISTOCOMPATIBILITY COMPLEX. MeSH, 1987

 a group of alleles in an organism that are inherited together from a single parent.[1][2] However, there are other uses of this term. First, it is used to mean a collection of specific alleles (that is, specific DNA sequences) in a cluster of tightly linked genes on a chromosome that are likely to be inherited together—that is, they are likely to be conserved as a sequence that survives the descent of many generations of reproduction.[3][4] A second use is to mean a set of linked single-nucleotide polymorphism (SNP) alleles that tend to always occur together (i.e., that are associated statistically). It is thought that identifying these statistical associations and few alleles of a specific haplotype sequence can facilitate identifying all other such polymorphic sites that are nearby on the chromosome. Such information is critical for investigating the genetics of common diseases; which in fact have been investigated in humans by the International HapMap Project.[5][6] Thirdly, many human genetic testing companies use the term in a third way: to refer to an individual collection of specific mutations within a given genetic segment; (see short tandem repeat mutation). Wikipedia accessed 2018 Nov 8 https://en.wikipedia.org/wiki/Haplotype

A haplotype is the set of SNP alleles along a region of a chromosome. Theoretically there could be many haplotypes in a chromosome region, but recent studies are typically finding only a few common haplotypes. Developing a Haplotype map of the human genome, 2001 http://www.genome.gov/10001665 

A particular pattern of sequential SNPs found on a single chromosome. These SNPs tend to be inherited together over time and can serve as disease-gene markers. The examination of single chromosome sets (haploid sets), as opposed to the usual chromosome pairings (diploid sets), is important because mutations in one copy of a chromosome pair can be masked by normal sequences present on the other copy. 

From “haploid genotype.”  The key idea is that alleles often travel together. Related terms: haplotyping, haplotyping technologies Cell biology diploid, haploid, ploidy; Maps & mapping: haplotype map HapMap; Narrower term: SNPs & genetic variations haploinsufficiency, haplotype block, SNP haplotype

haplotyping: 
Haplotyping involves grouping subjects by haplotypes, or particular patterns of sequential SNPs, found on a single chromosome. These SNPs tend to be inherited together over time and can serve as disease-gene markers.

Somatic cells, as opposed to germ cells, have two copies of each chromosome. A given single- base position may be homozygous for the wild- type base (each chromosome has the normal allele), homozygous for a SNP base (each chromosome has the altered allele), or heterozygous for two different bases (one chromosome has the normal allele and the other has the abnormal allele). Haplotyping involves grouping subjects by haplotypes, or particular patterns of sequential SNPs, found on a single chromosome. These SNPs tend to be inherited together over time and can serve as disease- gene markers. The examination of single chromosome sets (haploid sets), as opposed to the usual chromosome pairings (diploid sets), is important because mutations in one copy of a chromosome pair can be masked by normal sequences present on the other copy.  Genes tend to travel in packs. This is good news for pharmacogenomics. Broader terms genotyping, sequencing

haplotyping technologies: Include microarrays,   mass spectrometry,   sequencing

immunosequencing:  a platform technology that allows the enumeration, specification and quantification of each and every B-and/or T-cell in any biologic sample of interest. It is based on bias-controlled multiplex PCR and high throughput sequencing and is highly accurate, standardized, and sensitive.  Immune monitoring technology primer: immunosequencing, Ilan Kirsch J Immunother Cancer. 2015; 3: 29. Published online 2015 Jun 25. doi:  10.1186/s40425-015-0076-y  https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4480450/

Maxam-Gilbert sequencing & Sanger sequencing: a method of DNA sequencing developed by Allan Maxam and Walter Gilbert in 1976–1977. This method is based on nucleobase-specific partial chemical modification of DNA and subsequent cleavage of the DNA backbone at sites adjacent to the modified nucleotides.[1]   … Maxam–Gilbert sequencing was the first widely adopted method for DNA sequencing, and, along with the Sanger dideoxy method, represents the first generation of DNA sequencing methods. Maxam–Gilbert sequencing is no longer in widespread use, having been supplanted by next-generation sequencing methods. Wikipedia accessed 2018 March 16 https://en.wikipedia.org/wiki/Maxam%E2%80%93Gilbert_sequencing

microsequencing: Sequencing of proteins or peptides in very small amounts (sub microgram), sometimes for use as probes.

minisequencing: A solid- phase method for the detection of any known point mutation or allelic variation of DNA. In the method amplified, biotinylated DNA sequences containing the mutation site are immobilized onto streptavidin coated microplate and primer extension reactions are carried out using labeled nucleotides. Incorporation of the labeled nucleotide is dependent on the genotype and is analyzed using ELISA technique. Assay method allows automation. Photometry applications, Labsystems Oy, Finland, no longer on website

Single base sequencing. 

multilocus sequence typing: Direct nucleotide sequencing of gene fragments from multiple housekeeping genes for the purpose of phylogenetic analysis, organism identification, and typing of species, strain, serovar, or other distinguishable phylogenetic level. MeSH 2011

nanopore sequencing: a third generation[1] approach used in the sequencing of biopolymers- specifically, polynucleotides in the form of DNA or RNA. Wikipedia accessed 2018 Sept 4
https://en.wikipedia.org/wiki/Nanopore_sequencing
 

next generation sequencing: High-throughput (formerly "next-generation") sequencing applies to genome sequencing, genome resequencing, transcriptome profiling (RNA-Seq), DNA-protein interactions (ChIP-sequencing), and epigenome characterization.[55].  Resequencing is necessary, because the genome of a single individual of a species will not indicate all of the genome variations among other individuals of the same species.  The high demand for low-cost sequencing has driven the development of high-throughput sequencing technologies that parallelize the sequencing process, producing thousands or millions of sequences concurrently.[56][57][58] High-throughput sequencing technologies are intended to lower the cost of DNA sequencing beyond what is possible with standard dye-terminator methods.[59] In ultra-high-throughput sequencing as many as 500,000 sequencing-by-synthesis operations may be run in parallel.[60][61][62]  Wikipedia accessed 2018 March 16 https://en.wikipedia.org/wiki/DNA_sequencing#Next-generation_methods

Track 5: Next-Gen Sequencing Informatics
Next-Gen Sequencing Informatics Advances in Large-Scale Computing 2019 April 17-18 Boston MA  Program   Tremendous advancements have been made to broaden NGS applications from research to the clinic. Especially as genomics becomes more integrated with precision medicine initiatives. In spite of this, enormous challenges for NGS still exist including data analysis pipelines and platforms; data integration, interpretation and visualization; application of sequencing to cancer, immunology, diagnostics, and therapeutic development and emerging sequencing technologies

optical mapping: Stretching DNA molecules in nanochannels allows structural and copy-number variations to be visualized like beads on a string. Channeling DNA for optical mapping Yael Michael  & Yuval Ebenstein  Nature Biotechnology 30,  762–763 (2012)  doi:10.1038/nbt.2324 published online August

Next generation sequencing (NGS) is revolutionizing all fields of biological research but it fails to extract the full range of information associated with genetic material. Optical mapping of DNA grants access to genetic and epigenetic information on individual DNA molecules up to 1 Mbp in length.  Beyond sequencing: optical mapping of DNA in the age of nanotechnology and nanoscopy Michal Levy-Sakin,  Yuval Ebenstein  Current Opinion in Biotechnology Volume 24, Issue 4, August 2013, Pages 690–698  http://www.sciencedirect.com/science/article/pii/S0958166913000128

pathogen sequencing: In the future, more pathogens will have their genomes completely sequenced to determine not only how the pathogen causes disease, but what, if any, treatments will be most effective. The DNA sequences of viruses like HIV, human papilloma virus (HPV), and hepatitis C (HCV) are already being characterized and therapies prescribed based on this genetic information. To perform these types of diagnoses, DNA sequencing will have to become faster, more cost effective, simpler to perform, and more accessible to clinical laboratories. 

PCR and NGS-Based Molecular Diagnostics March 14-15, 2019 San Francisco, CA Program |   Advances Techniques and Tools for Precision Medicine Advances in molecular diagnostics technologies have sparked innovation, expanded research capabilities, and enhanced clinical diagnostics. Cambridge Healthtech Institute’s 6th Annual PCR and NGS-Based Diagnostics symposium puts an emphasis on the NGS and PCR technologies that drive precision medicine and showcases how they are being used to alter clinical outcomes. This event will provide a comprehensive look at integrating molecular diagnostics solutions for biomarker discovery and development, point-of-care, companion diagnostics, and infectious disease.

published working drafts - human genome: International Human Genome Sequencing Consortium special issue: Nature 409 (6822) 15 Feb 2001 
https://www.nature.com/nature/volumes/409/issues/6822

Human Genome [Celera Genomics sequence] special issue: Science 291 (5507) Feb. 16, 2001 http://www.sciencemag.org/content/vol291/issue5507/index.shtml

resequencing: Eric Lander, director of the Whitehead Institute's Center for Genome Research, and professor of biology at MIT notes " The human genome will need to be sequenced only once, but it will be resequenced thousands of times, in order, for example to unravel the polygenic factors underlying human susceptibilities and predispositions … Re-sequencing will also provide the ultimate tool for genotyping studies" E. Lander "The New Genomics" Science 274: 536, 25 Oct. 1996

Previously sequenced site is resequenced for SNP discovery or other purposes.  DNA resequencing involves sequencing a DNA region where a reference sequence for the region is already available. These studies provide important insight into the function of genes and the evolution of genes and populations. Applications abound including: comparative genomics, high-throughput SNP detection, identifying mutant genes in disease pathways, profiling transcriptomes for organisms where little information is available, researching lowly expressed genes, to identifying newly emerging or genetically engineered bacterial and viral strains. 

RNA-Seq (RNA sequencing):  also called whole transcriptome shotgun sequencing[2] (WTSS), uses next-generation sequencing(NGS) to reveal the presence and quantity of RNA in a biological sample at a given moment.[3][4] RNA-Seq is used to analyze the continuously changing cellular transcriptome. Wikipedia accessed 2018 Oct 21 https://en.wikipedia.org/wiki/RNA-Seq

Sanger sequencing
: See under Maxam-Gilbert sequencing.

scanning, scoring: SNPs & other genetic variations

sequence coverage: refers to the general concept of aiming for high number of unique reads of each region of a sequence.[3]  Wikipedia accessed 2018 Aug 28 

https://en.wikipedia.org/wiki/Coverage_(genetics)

sequence inversion: The deletion and reinsertion of a segment of a nucleic acid sequence in the same place, but flipped in an opposite orientation. MeSH 2010

sequencing: Proteins, nucleic acids -- Analytical procedures for the determination of the order of amino acids in a polypeptide chain or of nucleotides in a DNA or RNA molecule. IUPAC Compendium

Largely automated now. Full DNA sequencing is the "gold standard" for genotyping.   Narrower terms; next generation sequencing, shotgun sequence, de novo sequencing, microsequencing, minisequencing, multiplex sequencing, Sanger sequencing, sequencing by synthesis.  Related terms: genotyping, GWAS Genome Wide Association Sequencing, haplotyping, sequencing data analysis & storage, sequencing data management

sequencing by synthesis: Promising new sequencing technologies, based on sequencing by synthesis (SBS), are starting to deliver large amounts of DNA sequence at very low cost. Polymorphism detection is a key application. Quality scores and SNP detection in sequencing-by-synthesis systems., Brockman W, Alvarez P, Young S, Garber M, Giannoukos G, Lee WL, Russ C, Lander ES, Nusbaum C, Jaffe DB, Genome Research 2008 Jan 22

The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 https://bitesizebio.com/13546/sequencing-by-synthesis-explaining-the-illumina-sequencing-technology/  

sequencing - cost of: Cheap and easy genome sequencing has been both a blessing and a curse. We are able to find an incredible wealth of variation, but for the most part we have no easy way to tell whether a difference might contribute to a disease or not. The poster child for this problem is autism. Lots of genome wide association studies (GWAS) have been done and lots of rare variants in lots of different genes have been found – unfortunately, way too many to pick out the ones that really matter.  Luckily our friend yeast can help. Yeast winnows down GWAS hits in autism, SGD Database 2013 https://www.yeastgenome.org/blog/yeast-winnows-down-gwas-hits-in-autism  Related term: Molecular Diagnostics $1,000 genome

   Cost per raw megabase of DNA Sequence, NIH NHGRI 2001-2017

sequencing - high- throughput: Uses robotics, automated DNA- sequencing machines and computers.

shotgun sequencing: Sequencing method which involves randomly sequencing tiny cloned pieces of the genome, with no foreknowledge of where on a chromosome the piece originally came from. This can be contrasted with "directed" [sequencing] strategies, in which pieces of DNA from adjacent stretches of a chromosome are sequenced. Directed strategies eliminate the need for complex reassembly techniques. Because there are advantages to both strategies, researchers expect to use both random (or shotgun) and directed strategies in combination to sequence the human genome. DOE

Single-Cell Sequencing
Single-Cell Sequencing August 20-24 2018 • Washington, DC Program | 
The unthinkable is now possible. Next-generation sequencing (NGS) has evolved rapidly, reducing costs and making cancer genome sequencing more routine. Traditional approaches requiring bulk DNA or RNA from multiple cells only provide global information on average states of cell populations without resolving genomic differences in heterogeneous tumors. But now, whole-genome amplification (WGA) and NGS advances enable analyses of single cells to detect variations in individual cancer cells and dissect tumor evolution. Thus, single-cell sequencing will improve oncology by detecting rare tumor cells early, monitoring circulating tumor cells (CTCs), measuring intra-/intertumor heterogeneity, guiding chemotherapy and controlling drug resistance – all aiding cancer diagnosis, prognosis and prediction and leading to individualized cancer therapy.

third-generation sequencing (TGS): Sequencing single DNA molecules without the need to halt between read steps (whether enzymatic or otherwise).  A window into third-generation sequencing, Glossary Eric E. Schadt*, Steve Turner Andrew Kasarskis, Human Molecular Genetics 19, IssueR2 Pp. R227-R240. http://hmg.oxfordjournals.org/content/19/R2/R227

transcriptome sequencing: Deep sequencing of transcriptomes, also known as RNA-Seq, provides both the sequence and frequency of RNA molecules that are present at any particular time in a specific cell type, tissue or organ.[8] Counting the number of mRNAs that are encoded by individual genes provides an indicator of protein-coding potential, a major contributor to phenotype.[9] Improving methods for RNA sequencing is an active area of research both in terms of experimental and computational methods.[10]  Wikipedia accessed 2018 Aug 28 https://en.wikipedia.org/wiki/Coverage_(genetics)#Transcriptome_sequencing

ultra-deep sequencing:  
The term "ultra-deep" can sometimes also refer to higher coverage (>100-fold), which allows for detection of sequence variants in mixed populations.[4][5][6]In the extreme, error-corrected sequencing approaches such as Maximum-Depth Sequencing can make it so that coverage of a given region approaches the throughput of a sequencing machine, allowing coverages of >10^8.[7] Wikipedia accessed 2018 Aug 28 https://en.wikipedia.org/wiki/Coverage_(genetics)#Ultra-deep_sequencing

viral genotyping: Genomic data is enabling researchers to predict a patient's response to therapy based on the viral genotype for viral infections. HIV genotyping is an early example of how treatment decisions are made based on the genotype of the virus.

Whole Exome Sequencing  Techniques to determine the complete complement of sequences of all EXONS of an organism or individual. MeSH 2018

Whole Genome Sequencing: Techniques to determine the entire sequence of the GENOME of an organism or individual. Year introduced: 2018 MeSH

whole genome shotgun sequencing: Whole Genome Shotgun (WGS) sequencing projects are incomplete genomes or incomplete chromosomes that are being sequenced by a whole genome shotgun strategy. WGS projects may be annotated, but annotation is not required. The pieces of a WGS project are the contigs (overlapping reads), and they do not include any gaps. NCBI Whole Genome Shotgun Submissions http://www.ncbi.nlm.nih.gov/genbank/wgs.html   Broader term shotgun sequencing methodRelated term: GWAS Genome Wide Association Sequencing  Wikipedia http://en.wikipedia.org/wiki/Shotgun_sequencing#Whole_genome_shotgun_sequencing   

Sequencing resources
Ensembl Glossary http://uswest.ensembl.org/common/Help/Glossary?db=core 
European Nucleotide Archive  https://www.ebi.ac.uk/ena   provides a comprehensive record of the world's nucleotide sequencing information, covering raw sequencing data, sequence assembly information and functional annotation
IUPAC  International Union of Pure and Applied Chemistry, Glossary for Chemists of terms used in biotechnology. Recommendations, Pure & Applied Chemistry 64 (1): 143-168, 1992. 200 + definitions.
IUPAC International Union of Pure and Applied Chemistry, Glossary of Terms used in Bioinorganic Chemistry, Recommendations, 1997. 450+ definitions. http://www.chem.qmw.ac.uk/iupac/bioinorg/
NCBI (US) BLAST Glossary, 20011 http://www.ncbi.nlm.nih.gov/Education/BLASTinfo/glossary2.html
NHGRI (National Human Genome Research Institute), Talking Glossary of Genetic Terms, 100+ definitions. https://www.genome.gov/genetics-glossary  Includes extended audio definitions.

NCBI Sequence analysis https://www.ncbi.nlm.nih.gov/guide/sequence-analysis/

Technologies Conferences http://www.healthtech.com/conferences/upcoming.aspx?s=TCH
Technologies Short courses http://www.healthtech.com/Conferences_Upcoming_ShortCourses.aspx?s=TCH

How to look for other unfamiliar  terms

IUPAC definitions are reprinted with the permission of the International Union of Pure and Applied Chemistry.

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