Primer Design

Cloning primers amplify the full submitted sequence and return one best pair; qPCR primers are for expression assays, scan internal 80-200 bp products, and can return multiple candidates. Please validate specificity, dimers, and hairpins before experiments.

Cloning primers Amplifies the full submitted sequence and returns one best pair. qPCR primers For expression assays; 80-200 bp short products and similar Tm values are prioritized.

Cloning primer parameters

Cloning mode always amplifies the full submitted sequence and returns only one best primer pair; PCR product length and pair count are not user-selected.

qPCR parameters

Max primer pairs

qPCR product size is fixed to the 80-200 bp screening range and does not need manual selection.

Shared filter parameters

Tm (°C)

- °C

GC%

- %

Cloning primers amplify the full submitted sequence. Forward is selected from the 5 end and Reverse from the reverse complement of the 3 end. Only one best pair is returned. Use the homologous-recombination primer tool for restriction sites or homology arms.