{"uid":"cap_rPX4-MA_Fn6AgZqFLeM0n","slug":"qpcr-relative-expression-calculator-pfaffl-method-85dc7738","name":"qPCR Relative-Expression Calculator (Pfaffl Method)","description":"Calculate a Pfaffl expression ratio from supplied Ct values and amplification efficiencies; require a declared stable reference. No diagnostic interpretation. New modular adapter: retained result recovery for 24 hours, minimal receipt metadata for 90 days. Existing live route unchanged.","url":"https://www.mahastrategies.com/api/v1/compute/biology/qpcr-relative-expression?utm_source=zero.xyz","method":"POST","headers":{},"bodySchema":{"type":"object","properties":{"dataClass":{"enum":["public","synthetic"],"type":"string"},"targetSampleCt":{"type":"number","maximum":1000000000000,"minimum":-1000000000000},"referenceStable":{"type":"boolean"},"targetControlCt":{"type":"number","maximum":1000000000000,"minimum":-1000000000000},"targetEfficiency":{"type":"number","maximum":1000000000000,"minimum":-1000000000000},"referenceSampleCt":{"type":"number","maximum":1000000000000,"minimum":-1000000000000},"referenceControlCt":{"type":"number","maximum":1000000000000,"minimum":-1000000000000},"referenceEfficiency":{"type":"number","maximum":1000000000000,"minimum":-1000000000000}}},"responseSchema":null,"example":null,"exampleRequest":null,"tags":["x402"],"displayCostAmount":"0.010043","displayCostAsset":"USDC","priceDynamic":false,"priceHint":null,"priceStatus":"priced","priceSource":"probe","requiresHandshake":false,"reviewCount":0,"rating":{"score":"0.00","successRate":"0.00","reviews":0,"stars":null,"state":"unrated"},"availabilityStatus":"unknown","priceObserved":null,"sessionDeposit":null,"pricing":{"kind":"static","summary":"$0.010043/call","primary":{"kind":"static","protocol":"x402","network":"base","amountUsd":"0.010043","per":"call","confidence":"exact"},"accepted":[{"kind":"static","protocol":"x402","network":"base","amountUsd":"0.010043","per":"call","confidence":"exact"}]},"paymentMethods":[{"uid":"pm_P3IBvSXp6HjHGjrIRzFy1","protocol":"x402","methodType":"crypto","chain":"base","mode":"charge","costAmount":"0.010043","costPer":"request","priority":0,"asset":"0x833589fCD6eDb6E08f4c7C32D4f71b54bdA02913","unit":"request","depositMicros":null,"planRef":null}],"brandName":null,"brandSlug":null,"brandBaseUrl":null,"brandDocsUrl":null,"whatItDoes":"Calculates a Pfaffl expression ratio from target and reference gene Ct values and amplification efficiencies, requiring a declared stable reference gene.","exampleAgentPrompt":"Calculate the Pfaffl relative expression ratio for my qPCR experiment: target gene sample Ct is 22.3, target control Ct is 28.1, target efficiency is 1.95; reference gene sample Ct is 19.4, reference control Ct is 19.6, reference efficiency is 1.98 — the reference gene is confirmed stable, and this is synthetic data.","exampleUseCases":[{"title":"Housekeeping gene-normalized fold change","prompt":"I've run a qPCR experiment on treated vs untreated cells. Target gene sample Ct is 24.5, target control Ct is 30.2, target efficiency 1.92; GAPDH reference sample Ct is 20.1, reference control Ct is 20.3, reference efficiency 1.99. GAPDH is confirmed stable. Can you compute the Pfaffl expression ratio for me? This is synthetic data."},{"title":"Efficiency-corrected gene expression comparison","prompt":"Please compute the Pfaffl ratio for my RT-qPCR results: target Ct in sample is 18.7, in control 25.4, efficiency 1.85; beta-actin reference Ct in sample is 17.2, in control 17.5, efficiency 1.97 — reference is stable and data class is public."},{"title":"Automated qPCR pipeline normalization step","prompt":"As part of my analysis pipeline, calculate the relative expression ratio using the Pfaffl method: target sample Ct 26.8, target control Ct 32.1, target efficiency 1.90; reference sample Ct 21.0, reference control Ct 21.3, reference efficiency 2.00. Mark reference as stable, data class synthetic."}],"resultDescription":"Returns a computed Pfaffl expression ratio derived from the supplied Ct values and amplification efficiencies for both target and reference genes. Results are retained for 24 hours; minimal receipt metadata is retained for 90 days. No diagnostic interpretation is provided.","failureModes":["Missing or false referenceStable flag — endpoint requires reference gene to be declared stable","Out-of-range or invalid Ct/efficiency values outside expected biological bounds","Missing required fields such as targetSampleCt, referenceControlCt, or efficiencies","Invalid dataClass enum value (must be 'public' or 'synthetic')","Payment failure via x402 protocol preventing request processing","Server error or timeout during computation"],"whenToPreferThis":"Use this endpoint when you need a programmatic, efficiency-corrected Pfaffl relative expression ratio from raw Ct values without manual spreadsheet calculations. Prefer it over delta-delta Ct approaches when amplification efficiencies differ from 100% and must be accounted for. Best suited for automated qPCR analysis pipelines where a stable reference gene is already confirmed.","instructions":null,"reviewSummary":null,"reviewSummaryHighlights":null,"reviewSummaryConcerns":null,"reviewSummaryGeneratedAt":null,"activationCount":0,"lastUsedAt":null,"lastSuccessfullyRanAt":null,"lastHealthCheckAt":"2026-10-15T06:39:12.276Z","isFirstParty":false,"canonicalSlug":"qpcr-relative-expression-calculator-pfaffl-method-85dc7738"}