LAMP Primer Design Guideline

The essence of LAMP (Loop-mediated Isothermal Amplification) lies in its multi-primer system. Well-designed primers ensure both sensitivity and specificity, making primer design the key to successful amplification.

1. Primer Set

A standard LAMP assay requires 4 - 6 primers, recognizing 8 regions of the target sequence: 

• Outer primers（F3/B3）
• Inner primers（FIP/BIP）
• Loop primers（LF/LB，optional）

2. Target Sequence Selection

• Length
• Specificity
• Avoid secondary structures

3. Key Primer Design Parameters

Length, Melting Temp (Tm), GC Content, 3'-End Stability, Avoid Dimers

4. Loop Primer (LF/LB) Design (Optional)

• Bind to the Fc or Bc regions (between F1/B1 and F2/B2), typically 18–22 nt in length.
• Tm ≈ 60-65℃, working synergistically with inner primers to boost amplification efficiency.

5. Recommended Tools

• Primer design software：https://lamp.neb.com/#!/
• Specificity check：NCBI BLAST

6. Validation and Optimization

• Specificity test: Use BLAST to rule out off-target binding.
• Sensitivity test: Perform template serial dilutions to determine the limit of detection (LOD).
• Colorimetric compatibility: For assays using dyes, ensure primers do not interfere with colorimetric signals.

Appendix: EZassay Biotech™LAMP Product Portfolio

For product information and technical support, please contact EZassay Biotech™.

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