Directionality: leading and lagging strands

Molecules (Continuity and change) · DNA replication · note 5 of 6

Spec D1.1.6, D1.1.7
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Directionality: leading and lagging strandsSpec D1.1.6, D1.1.7

In short

DNA polymerases only add the 5' end of a new nucleotide to the 3' end of a growing strand, so new DNA is built in the 5' to 3' direction. At a replication fork the leading strand is therefore made continuously, from one RNA primer, while the lagging strand is made discontinuously as Okazaki fragments, each needing its own primer.

5' and 3' ends

Each nucleotide in a strand is linked to the next through its deoxyribose. The 5' terminal of a strand has a phosphate group attached to carbon 5' of the sugar; the 3' terminal has a free –OH group on carbon 3'. The two strands of the double helix are antiparallel: they run in opposite directions.

DNA polymerases add the 5' of a DNA nucleotide to the 3' end of a strand of nucleotides. The phosphate on carbon 5' of the incoming nucleotide is bonded to the 3' –OH of the strand. Free nucleotides arrive as deoxynucleoside triphosphates, and the release of two phosphates provides the energy for the bond. New DNA therefore always grows in the 5' → 3' direction.

Leading and lagging strands

Replication at a replication fork
FeatureLeading strandLagging strand
Direction of synthesis relative to the forkSame direction as the fork movesOpposite direction to the fork
Type of replicationContinuousDiscontinuous, in short sections called Okazaki fragments
RNA primersNeeded only once, at the startNeeded repeatedly, one for each Okazaki fragment
JoiningLittle joining: the single primer is replaced by DNA and the one nick sealed by DNA ligaseFragments joined by DNA ligase

Because the template strands are antiparallel and synthesis can only go 5' → 3', one new strand can follow the helicase as the fork opens, while the other must be made in short pieces backwards from the fork, each started afresh as more template is exposed.

Replication fork in a prokaryote: helicase at the fork, parental strands labelled 3′ and 5′; the leading strand is made continuously towards the fork from one RNA primer by DNA polymerase III; the lagging strand is made away from the fork as Okazaki fragments, with DNA primase making RNA primers, DNA polymerase III extending fragments, DNA polymerase I replacing primers and DNA ligase sealing the gaps; 5′ and 3′ ends are marked on the new strands. (opens full size in a new tab)
At a replication fork, new DNA always grows 5′ → 3′: the leading strand is continuous and the lagging strand is made in Okazaki fragments.
Exam tip:

Linking question: what biological mechanisms rely on directionality? Replication, transcription and translation all proceed 5' to 3', which follows from the structure of the sugar–phosphate backbone.

Written and checked against the IB Biology HL specification · Updated October 2026

Frequently asked questions

Why is DNA replication semi-conservative?

DNA replication is semi-conservative because the two strands of the original molecule separate and each acts as a template for a new strand. Each daughter molecule therefore keeps one original strand and gains one new strand. Complementary base pairing makes the new strand match the old partner, so both copies have identical base sequences.

What are the three stages of PCR?

The three stages of PCR are denaturation, annealing and extension. At about 95 °C hydrogen bonds break and the strands separate; at about 55 °C primers bind to the ends of the target sequence; at about 72 °C heat-stable Taq polymerase builds new strands. Each cycle doubles the number of copies.

How does gel electrophoresis separate DNA fragments?

Gel electrophoresis separates DNA fragments by length. DNA is negatively charged because of its phosphate groups, so when a voltage is applied it moves through the gel towards the positive electrode. Smaller fragments move through the gel mesh more easily, so they travel further than larger fragments in the same time.

All 5 questions on DNA replication