Skatebro:Primer Design

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(Tips)
Line 2: Line 2:
==Tips==
==Tips==
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*In general, want highest GC content possible
+
*In general, want high GC content, up to 50%
*Insert a double stop codon (taa taa)
*Insert a double stop codon (taa taa)
-
*Binding length of 18-22 bps is ideal plus tails
+
*Binding length of 18-22 bps is ideal
-
*Melt temps. of forward and reverse primers should be within ~2c and ~50c is desirable
+
*Melt temps. of forward and reverse primers should be within ~2c and ~60c is desirable
 +
*Make sure primer can only form short/thermodynamically unstable hairpins

Revision as of 03:53, 7 February 2008

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 ~60c is desirable
  • Make sure primer can only form short/thermodynamically unstable hairpins
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