Since its launch in 2009, the MFEprimer server has been continuously operating for 16 years. We remain committed to maintaining this platform as a sustainable, long-term resource for the global research community.

From version 4.0, the MFEprimer Web Server serves as a comprehensive platform for PCR primer quality control and design tools. We have recently launched a new feature, the MP-Ref, which is a multiplex PCR primer design tool based on reference genome. Additionally, we are developing a primer design project aimed at diverse genomes. Below is a detailed list of our tools:

Quality Control Tools

• Specificity Check: This tool allows for the verification of primer specificity across multiple background databases, ensuring minimal off-target effects.
• Dimer Check: This tool assesses your primers for potential dimer formations, which can inhibit PCR performance.
• Hairpin Check: Evaluate primers for hairpin structures that can negatively impact PCR efficiency.
• Coverage Analysis for Microbes: This tool assesses primer coverage for specific microbial strains, helping to identify potential primer issues.

Primer Design Tools

• MP-Ref: Multiplex PCR Primer Deisgn for Reference Genomes. Our enhanced multiplex PCR primer design tool accommodates a wide range of applications, streamlining the design process for complex PCR setups.

History

• 2019, MFEprimer-3.0: quality control for PCR primers.
• 2012, MFEprimer-2.0: a fast thermodynamics-based program for checking PCR primer specificity.
• 2009, MFEprimer-1.0: multiple factor evaluation of the specificity of PCR primers.