How do I choose primers for Sanger sequencing?

Choosing the right primers is a crucial factor in successful automated Sanger DNA sequencing. Here are a few things to keep in mind when choosing primers for Sanger sequencing: 

• Length of the primer should be in the range of 18 and 24 bases. 
• The primer must have a GC content of about 45-55%. 
• The primer must have a GC-lock on the 3' end. 
• The melting temperature of the primer should be higher than 50°C but lower than 65°C. 
• The primer should be located at least 50-60 bases upstream of your sequence of interest.
• The primer should not include poly base regions. 
• ·Check primer for specificity in annealing to template. 
• Avoid primers with secondary structures or the potential to self-hybridize.