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PCR Annealing Temperature Calculator (Ta = Tm − 5°C)

PCR Ta by the simple rule (lower primer Tm − 5°C) or the Rychlik 1990 formula (0.3·Tm_primer + 0.7·Tm_product − 14.9), with a ±3°C gradient window and primer-Tm mismatch warning. Derived from Rychlik 1990, Innis & Gelfand 1990, Lorenz 2012. Lab-planning use only.

json· JSON resultOpen tool

Quick start

Call this tool from your code in three languages.

cURL

curl -X POST 'http://127.0.0.1:3003/en/api/tools/annealing-temperature-calculator' \
  -H 'Content-Type: application/json' \
  -d '{"rule":"simple","forwardTm":62,"reverseTm":60,"productTm":85,"decimalPlaces":1}'

API reference

Send a POST request with your inputs as JSON. File parameters require a separate upload first.

Endpoint

HTTP

POST http://127.0.0.1:3003/en/api/tools/annealing-temperature-calculator

Parameters

Name	Type	Required	Description
rule	select	Yes	Simple rule uses the lower primer Tm; Rychlik also weighs the product Tm.
forwardTm	number	Yes	Melting temperature of the forward primer.
reverseTm	number	Yes	Melting temperature of the reverse primer.
productTm	number	Yes	Amplicon melting temperature (typically 75–90°C for a 100–500 bp product); used by the Rychlik formula.
decimalPlaces	number

MCP integration

Add this tool to your Model Context Protocol server so AI agents can list and call it.

Server configuration

Add this block to your MCP client configuration:

mcp.json

{
  "mcpServers": {
    "elysiatools-annealing-temperature-calculator": {
      "name": "annealing-temperature-calculator",
      "description": "PCR Ta by the simple rule (lower primer Tm − 5°C) or the Rychlik 1990 formula (0.3·Tm_primer + 0.7·Tm_product − 14.9), with a ±3°C gradient window and primer-Tm mismatch warning. Derived from Rychlik 1990, Innis & Gelfand 1990, Lorenz 2012. Lab-planning use only.",
      "baseUrl": "http://127.0.0.1:3003/mcp/sse?toolId=annealing-temperature-calculator",
      "command": "",
      "args": [],
      "env": {},
      "isActive": true,
      "type": "sse"
    }
  }
}

List available tools

After connecting to the SSE endpoint, list the exposed tools:

tools/list

{
  "jsonrpc": "2.0",
  "id": 1,
  "method": "tools/list"
}

Call this tool

Invoke the tool by its id, passing arguments built from its parameters:

tools/call

{
  "jsonrpc": "2.0",
  "id": 2,
  "method": "tools/call",
  "params": {
    "name": "annealing-temperature-calculator",
    "arguments": {
  "rule": "simple",
  "forwardTm": 62,
  "reverseTm": 60,
  "productTm": 85,
  "decimalPlaces": 1
}
  }
}

Chain multiple tools in one session with a comma-separated toolId list, e.g. /mcp/sse?toolId=png-to-webp,jpg-to-webp,gif-to-webp (max 20).

Questions or issues? Contact [email protected]

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