Automated DNA Sequencing - Primers
Good primer design is one of the key factors in obtaining sound
sequencing data. Using either online tools or stand-alone programs,
please design your primers according to the recommended properties
below. Shown below is a small list of freely available tools for primer
design. Please DO NOT add two PCR primers into your reaction. One
sequencing reaction, one primer please!
Primer Design Tools
Recommended primer properties
- Primer concentration = 5µM.
- 18-26 bases long.
- Optimum Tm is great than 50 oC but less than 65oC.
- Around 50% GC.
- Avoid primers with secondary structures or the potential to
- Avoid runs of a single base (e.g. 3 to 4 repeats of the same
UMass User can order from Fisher Scientific for free shipping - Click the
Free Primers Provided by Umass Genomics Resource Lab