Amplify
qPCR Design
Design a qPCR assay against a region of an open sequence, then analyse the data that comes back from the instrument. Design and analysis sit in the same tool, so the assay a result belongs to is the assay you designed rather than a row you re-typed.
The real qPCR corpus: ΔΔCt (separate + same-tube), a standard curve, a significance table, Vandesompele 2002 geNorm in wide and long form, real amplification curves. Opens on the human GAPDH and MYC transcripts those experiments actually measure, plus a 72 bp miRNA precursor that walks Design right up to its length limit
What it does
- 01TaqMan primer and probe design over a selected region or a whole sequence
- 02Designed primers and probe written back onto the map as annotations
- 03Relative quantification by the delta-delta Ct method
- 04Amplification curve analysis, converting a curve to a Cq value
- 05Reference-gene stability ranking for normalisation
- 06Worked example datasets for each analysis tab, from published data rather than invented numbers
What it runs on
Primer thermodynamics come from the nearest-neighbour Tm engine in packages/ports; the analysis methods follow their published definitions. The panel runs on @ports/discover, @ports/eds, @ports/melt, @ports/qpcr, @ports/rdml, @ports/workflow.
Everything runs client-side. Files are parsed in the browser and projects are saved on this machine, so an unpublished sequence is never uploaded.
Questions
- Can I analyse Ct values without designing an assay first?
- Yes. The analysis tabs accept pasted Ct data directly, and each one ships a real example dataset so the expected input shape is visible.
- Does it rank reference genes?
- Yes. Candidate reference genes are ranked by expression stability, which is what decides whether a normaliser is usable in a given experiment.
Also searched as
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