PARAMETERS

OPTIMUM VALUES

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2.  G/C Clamp at the 3' End

3.  No self-complementarity

4.  No primer dimers (no complementarity to other primer)

5.  Random base distribution and composition

6.  Primer  Length

7.  Matched primer T_ms

8.  Distance and composition of amplified sequence

1-2 G/C (S) nucleotides             

< 3 contiguous bases

< 3 contiguous bases

45-55% G+C content

18-25 bases

100-600 bases apart