Custom multiplex tiling PCR panel design tool

Design tiling PCR primers to achieve complete coverage of the target sequences. Tiling PCR involves dividing a long genomic region into a series of short/long, overlapping amplicons, each of which is amplified by a dedicated primer pair. Adjacent amplicons share a defined overlap to ensure there are no gaps in the coverage. This makes the approach ideal for the high-throughput sequencing of large genes, the validation of variants across entire coding regions and the targeted resequencing of regions.

Retrieve sequence from NCBI by accession — chromosomes (NC_), mRNA (NM_), RefSeq contigs

NCBI Accession:

Upload local FASTA file — plain FASTA, multi-FASTA, or sequences

Upload FASTA:

Primer Design Options

Length range (nt)

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Tm range (°C)

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3′-end pattern

PCR product size (bp)

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Gap between amplicons

Non-specific priming control

Overlapping primers

C→T bisulfite conversion

Forward primer tail (5′-3′)

Reverse primer tail (5′-3′)

Need details? Sequence markup, parameters, output columns and the suggested workflow are documented in Help: PCR tool · Help: Troubleshooting
Exporting results: click inside the result area → Ctrl+A → Ctrl+C → paste into Excel.