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A base pair is an association of two nucleobases through hydrogen bonds in a compatible arrangement. It commonly joins opposite strands of a nucleic acid double helix, but can also join two regions of one folded RNA strand. Canonical Watson–Crick DNA base pairs match adenine with thymine and guanine with cytosine; canonical RNA base pairs use adenine with uracil instead. The nucleobases belong to complete nucleotides, including their attached sugars and backbone phosphate. See NHGRI: base pair.

The abbreviation bp also measures the length of a paired region. Our ACGTAC/TGCATG duplex has six base pairs, containing twelve nucleotides altogether. Its upper strand contains two adenine, two cytosine, one guanine, and one thymine nucleobase; the lower contains one, one, two, and two respectively. Combined, the counts are three of each nucleobase. Equality of total adenine and thymine, or of total guanine and cytosine, need not hold within either individual strand.

Base pairing is the recognition and association of compatible nucleobases. In canonical Watson–Crick geometry, an adenine–thymine base pair has two hydrogen bonds, while a guanine–cytosine base pair has three. These counts refer to those specific geometries and usual chemical forms; they are not a definition of every possible base pair. RNA can also contain other pairings, and canonical adenine–uracil pairing replaces adenine–thymine pairing. See Clark, Douglas and Choi, Biology 2e: DNA structure.

For this illustrative duplex, the six base pairs are A–T, C–G, G–C, T–A, A–T, and C–G. Thus three contribute two hydrogen bonds each and three contribute three each:Each dotted connector in the diagram counts one hydrogen bond; coloured boxes denote whole nucleobases, not individual chemical atoms. The solid rails denote the covalent bond scaffold. Separating the two strands disrupts base pairing and base stacking without requiring the backbone to be cut.

The count of fifteen hydrogen bonds is not fifteen identical contributions to a duplex's Gibbs free energy. Base stacking, interactions with water and ions, temperature, and the order of neighbouring base pairs also matter. Knowing the fraction of guanine–cytosine base pairs alone does not fix a nucleic acid double helix's shape or stability. See Brown, Genomes: pairing and stacking interactions. What makes base pairing especially useful is its selectivity: an exposed DNA strand constrains the partner's nucleic acid sequence, giving DNA replication a chemical recognition rule.

A deoxynucleoside triphosphate is a nucleotide containing deoxyribose and three phosphate groups. DNA polymerases use these substrates to extend DNA; the four standard substrate types carry adenine, cytosine, guanine or thymine.

Deoxyribonucleic acid, or DNA, is a nucleotide polymer that stores biological information in its nucleotide sequence. The crucial variable is order: two DNA molecules can contain the same numbers of each nucleobase yet carry different nucleotide sequences. This makes a physical molecule an inheritable record, rather than merely a mixture of ingredients. Genes are functional regions of that record; the whole DNA nucleotide sequence is not one long gene. See NHGRI: DNA fact sheet.

Each DNA strand has a backbone of alternating deoxyribose sugars and phosphate groups joined through covalent bonds. Attached nucleobases supply the familiar four-letter alphabet: adenine A, thymine T, guanine G, and cytosine C. In ordinary double-stranded DNA, base pairing matches A with T and G with C. The backbone connects successive nucleotides along a strand; base pairing connects partners across strands. Keeping these two connections distinct explains how strands can separate while each retains its own nucleotide sequence.

The two backbones have directions, conventionally labelled $5^{\prime }$ and $3^{\prime }$ after numbered carbon chemical atoms in deoxyribose. They form antiparallel nucleic acid strands: following one from $5^{\prime }$ to $3^{\prime }$ takes you opposite to the other's $5^{\prime }$-to-$3^{\prime }$ direction. For the chosen DNA nucleotide sequence $5^{\prime }\text{-}\mathrm{ATGCC}\text{-}{3}^{\prime }$, write its partner directly underneath as $3^{\prime }\text{-}\mathrm{TACGG}\text{-}{5}^{\prime }$. Now read that partner from its own $5^{\prime }$ end: its reverse complement is $5^{\prime }\text{-}\mathrm{GGCAT}\text{-}{3}^{\prime }$. Complementing letters and reversing their order are separate operations. See Alberts et al., Molecular Biology of the Cell: the structure and function of DNA.

The diagram's rails represent sugar–phosphate backbones and its coloured blocks represent nucleobases, not individual chemical atoms. Reading left to right gives five matched positions: A–T, T–A, G–C, C–G, C–G. There are two A–T-type base pairs and three G–C-type base pairs; the complete two-strand example contains ten nucleotides. Reversing the bottom row changes the reading convention, not the physical partners. Real double-stranded DNA winds into a nucleic acid double helix; the flattened ladder emphasizes base pairing and direction without pretending to show its three-dimensional geometry.

The motivating consequence is copying by a template. After strands separate, each existing DNA strand specifies which incoming nucleotides should accompany it through base pairing. DNA replication uses enzymes to build complementary strands, allowing the record to persist through successive biological cells. Base pairing supplies a selection rule, while enzymes perform the chemistry: a matching diagram alone is not a complete copying mechanism. The connection between structure and copying was already central to the model published by Watson and Crick on 25 April 1953. See Watson and Crick, Molecular Structure of Nucleic Acids, Nature 171, 737–738 (1953).

Stored information needs interpretation. Some DNA regions specify proteins through RNA; others contribute to functional RNA or regulate gene expression, and some have no known function. Non-coding DNA means sequence that does not specify a protein's sequence of amino acids, not automatically useless sequence. Nor does a DNA nucleotide sequence alone determine an organism's fate: phenotypes also depend on biological context and environment. See NHGRI: non-coding DNA. Its double-stranded form is often modeled mechanically as a worm-like chain, with elasticity and effective excluded volume depending on solvent conditions.

A nucleic acid sequence records the ordered nucleobases of a nucleic acid strand, conventionally read from $5^{\prime }$ to $3^{\prime }$. The standard DNA alphabet is adenine A, cytosine C, guanine G, and thymine T; the standard RNA alphabet uses uracil U in place of thymine T. Each letter denotes the nucleobase attached to one nucleotide, so the six-letter DNA nucleic acid sequence $5^{\prime }\text{-}\mathrm{ACGTAC}\text{-}{3}^{\prime }$ describes six linked nucleotides. See NHGRI: representing DNA by a sequence of letters.

Order carries information that composition loses. Both ACGTAC and AACCGT contain two adenine, two cytosine, one guanine, and one thymine nucleobase, but their nucleic acid sequences differ at positions 2, 3, 4, 5, and 6. At position 2, for example, the first has cytosine and the second adenine. Reporting only the four counts cannot reconstruct either ordered record.

To obtain the partner of $5^{\prime }\text{-}\mathrm{ACGTAC}\text{-}{3}^{\prime }$, apply the DNA base pairing rules position by position: A selects T, C selects G, G selects C, T selects A, A selects T, and C selects G. Alignment gives $3^{\prime }\text{-}\mathrm{TGCATG}\text{-}{5}^{\prime }$. Its opposite end labels matter because the two are antiparallel nucleic acid strands. Reading the partner from its own $5^{\prime }$ end reverses the aligned lettering and yields the reverse complement, $5^{\prime }\text{-}\mathrm{GTACGT}\text{-}{3}^{\prime }$.

Reversal alone gives CATGCA; complementation alone gives aligned TGCATG. Neither operation alone gives the partner written in the usual $5^{\prime }$-to-$3^{\prime }$ convention. Applying the reverse complement operation twice returns ACGTAC: GTACGT complements to CATGCA, which reverses to ACGTAC. This convention makes a single reported strand sufficient to specify an ideally complementary DNA duplex, provided its direction is known.