{"uid":"cap_IpwzbRrsjglZ0tx9kNvWN","slug":"qpcr-relative-expression-calculator-pfaffl-method-2c229b94","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.","url":"https://www.mahastrategies.com/api/v1/micro/biology-qpcr-relative-expression?utm_source=zero.xyz","method":"POST","headers":{},"bodySchema":{"type":"object","$schema":"https://json-schema.org/draft/2020-12/schema","required":["input"],"properties":{"input":{"type":"object","required":["type","method","bodyType","body"],"properties":{"body":{"type":"object","required":["dataClass","targetSampleCt","targetControlCt","referenceSampleCt","referenceControlCt","targetEfficiency","referenceEfficiency","referenceStable"],"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}},"additionalProperties":false},"type":{"type":"string","const":"http"},"method":{"enum":["POST"],"type":"string"},"bodyType":{"enum":["json","form-data","text"],"type":"string"}},"additionalProperties":false},"output":{"type":"object","required":["type"],"properties":{"type":{"type":"string"},"example":{"type":"object","required":["version","offerId","amountBaseUnits","inputDigest","result","boundaries","receiptDigest"],"properties":{"result":{"type":"object"},"offerId":{"enum":["biology-qpcr-relative-expression"],"type":"string"},"version":{"enum":["maha-microproducts/0.1"],"type":"string"},"boundaries":{"type":"array","maxItems":128,"minItems":0},"inputDigest":{"type":"string","maxLength":71},"receiptDigest":{"type":"string","maxLength":71},"amountBaseUnits":{"enum":["58001"],"type":"string"}},"additionalProperties":false}}}}},"responseSchema":null,"example":null,"exampleRequest":null,"tags":["x402"],"displayCostAmount":"0.058001","displayCostAsset":"USDC","priceDynamic":false,"priceHint":null,"priceStatus":"priced","priceSource":"registry","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":"$0.058001/call","primary":{"kind":"static","protocol":"x402","network":"base","amountUsd":"0.058001","per":"call","confidence":"exact"},"accepted":[{"kind":"static","protocol":"x402","network":"base","amountUsd":"0.058001","per":"call","confidence":"exact"}]},"paymentMethods":[{"uid":"pm_nm5j6HfD7n1Ps5rm28U2Y","protocol":"x402","methodType":"crypto","chain":"base","mode":"charge","costAmount":"0.058001","costPer":"request","priority":0,"asset":null,"unit":"request","depositMicros":null,"planRef":null}],"brandName":null,"brandSlug":null,"brandBaseUrl":null,"brandDocsUrl":null,"whatItDoes":"Calculates Pfaffl relative expression ratios from supplied Ct values and amplification efficiencies for target and reference genes, requiring a declared stable reference gene.","exampleAgentPrompt":"Calculate the Pfaffl relative expression ratio for my qPCR data: target sample Ct is 22.4, target control Ct is 25.1, reference sample Ct is 18.6, reference control Ct is 18.9, target efficiency is 1.95, reference efficiency is 1.98, and I confirm the reference gene is stable. Data class is synthetic.","exampleUseCases":[{"title":"Gene knockdown validation in treated cells","prompt":"I need to quantify how much my target gene is knocked down after siRNA treatment. Target sample Ct is 28.3, target control Ct is 22.1, reference sample Ct is 19.4, reference control Ct is 19.2, target PCR efficiency is 1.92, reference efficiency is 1.97 — the reference gene is confirmed stable. Data class is synthetic."},{"title":"Drug-induced gene upregulation measurement","prompt":"Can you compute the Pfaffl expression ratio to see how much a drug upregulates my gene of interest? Target sample Ct 20.5, target control Ct 26.8, reference sample Ct 21.0, reference control Ct 21.3, target efficiency 1.89, reference efficiency 1.96, reference is stable, data class public."},{"title":"Housekeeping gene normalization check","prompt":"Run a Pfaffl ratio calculation to compare expression levels between my treated and untreated samples: target sample Ct 24.7, control Ct 24.9, reference sample Ct 20.1, reference control Ct 20.0, target efficiency 1.94, reference efficiency 1.99, reference stable confirmed, data is synthetic."}],"resultDescription":"Returns a computed Pfaffl relative expression ratio result object, along with metadata including the offer ID (biology-qpcr-relative-expression), version, input digest, receipt digest, amount in base units, and any boundaries. No diagnostic interpretation is provided — only the numerical expression ratio.","failureModes":["Missing required Ct values or efficiencies returns a validation error","referenceStable set to false may be rejected or flagged as invalid input","Ct values or efficiencies outside expected biological range may still be accepted due to wide numeric limits but produce nonsensical ratios","Payment failure via x402 protocol prevents computation","Malformed body type or non-JSON content type causes request rejection"],"whenToPreferThis":"Choose this endpoint when you need a mathematically rigorous Pfaffl-corrected relative expression ratio that accounts for actual PCR amplification efficiencies rather than assuming 100% efficiency (as in the standard ΔΔCt method). Ideal when primer efficiencies differ between target and reference genes, or when high accuracy is required in quantitative expression analysis. Not suitable if you need diagnostic interpretation, absolute quantification, or multi-gene normalization across multiple reference genes simultaneously.","instructions":null,"reviewSummary":null,"reviewSummaryHighlights":null,"reviewSummaryConcerns":null,"reviewSummaryGeneratedAt":null,"activationCount":0,"lastUsedAt":null,"lastSuccessfullyRanAt":null,"lastHealthCheckAt":"2026-10-17T06:54:18.656Z","isFirstParty":false,"canonicalSlug":"qpcr-relative-expression-calculator-pfaffl-method-2c229b94"}