Digital PCR Primer Probe Design——Gene-π Class 2

In the last lecture, we introduced you to the principles and applications of digital PCR.

The design requirements of digital PCR primers and ordinary PCR primers are different: the purpose of ordinary PCR is to obtain the target gene, and the requirements for non-specific reactions and primer dimers are not strict; and digital PCR primers are the same as real-time PCR primers for non-specific reactions and primers The dimer requirements are strict

General principles of primer and probe design

▪ Choose the probe first, and then design the primers to be as close as possible to the probe

Basic principles of primer design:

▪ Amplified fragment length: 50-150 bases

Basic principles of probe design:

▪ Probe length: 13-25 bases (MGB probe)

Probe fluorescent label selection:

Principles of multiple probe design:

▪ The Tm values ​​of the primers or probes of all target genes should be similar.

Recommended primer probe design software: Beacon designer, Primer 3, Primer express, etc.

Join the GENE-π Digital PCR Technology School and swim in the ocean of digital PCR technology

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