DNA Structure and Properties Guide

DNA Base Pairing Rules

DNA follows Watson-Crick base pairing: Adenine (A) pairs with Thymine (T) via 2 hydrogen bonds. Guanine (G) pairs with Cytosine (C) via 3 hydrogen bonds. In RNA, Uracil (U) replaces Thymine. Chargaff's rules: in any double-stranded DNA, %A = %T and %G = %C. If GC content is 60%, then G = C = 30% each, A = T = 20% each. The human genome has approximately 40% GC content. Highly expressed genes tend to have higher GC content. Bacteria in extreme environments often have very high (>70%) or very low

DNA Melting Temperature

The melting temperature (Tm) is the temperature at which 50% of double-stranded DNA is single-stranded. G-C pairs have 3 hydrogen bonds (stronger than A-T's 2), so higher GC content raises Tm. Simple approximation: Tm = 4(G+C) + 2(A+T) in °C (for short oligonucleotides). For longer sequences with salt correction: Tm = 81.5 + 16.6 × log[Na⁺] + 0.41 × (%GC) − 675/length. Applications: PCR primer design requires primers with Tm in a similar range (typically 55-65°C). Hybridisation probes must be de

DNA Molecular Weight

Average molecular weight per base pair (dsDNA): approximately 650 Da (daltons) or 0.65 kDa. For dsDNA: MW ≈ 650 × bp (g/mol). More precisely: the four deoxyribonucleotide monophosphates have weights of 308 (dA), 332 (dC), 347 (dG), 323 (dT). Sodium salt form (common in molecular biology): add 22 Da per residue for Na⁺. Typical molecular biology sizes: PCR product 500 bp → 325 kDa. Gene 3,000 bp → 1.95 MDa. Human chromosome (average) → 150 MDa. E. coli genome 4.6 Mb → 3 GDa.

GC Content and Biology

GC content has important biological effects: gene expression: promoters and regulatory regions often have distinctive GC patterns. GC-rich regions form more stable secondary structures in RNA. Methylation: CpG dinucleotides (CG sequences) are methylation sites in mammals — CpG islands near gene promoters are often unmethylated in active genes. Codon usage: organisms with high GC content use different codons to code for the same amino acid (codon degeneracy allows this). Evolution: mutational bia

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