WangLab:Primer Design

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Primer design concerns

Primer concerns:

  1. GC>50%, Tm~66-72oC, both primers should have similar GC percentile and Tm.
  2. deltaG (free energy) for hairpin and dimmer close to Zero.


  • miniprep, purify with 30 ul water. Take 6 ul, digest , purify with 30 ul water. For subcloning, add 1 ul vector