Skatebro:Primer Design: Difference between revisions

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[[Skatebro:Project08]]


==Tips==
*In general, want high GC content, up to 50%
*Insert a double stop codon (taa taa)
*Binding length of 18-22 bps is ideal
*Melt temps. of forward and reverse primers should be within ~2c and ~50-60c is desirable
**analyze the landing sequence and the landing+tail Tm (can gradually increase annealing temp over pcr cycles if necessary)
*Make sure primer can only form short/thermodynamically unstable hairpins

Latest revision as of 03:08, 9 February 2008