What Factors Should Be Considered in Primer Design?
Several factors must be considered when designing primers for histological applications:
Specificity: Primers should bind only to the target DNA sequence to avoid non-specific amplification. Melting Temperature (Tm): The Tm of the primers should be around 55-65°C to ensure efficient binding during PCR. GC Content: Primers should have a GC content of 40-60% for stable binding. Primer Length: Typically, primers are 18-25 nucleotides long to balance specificity and binding efficiency. Avoiding Secondary Structures: Primers should not form hairpins, dimers, or other secondary structures that can interfere with the PCR process.
