Primer design

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1. PCR primer design

• Local software
Primer premier5, Oligo 6.1
• Web-based design
Primer3
(Whitehead Institute/MIT Center for Genome
Research)

Primer premier5

Principles for primer design
• Base composition
• Length
• Repeated and self-complementary seq.
• Cross dimer
• Tm
• 3’ Termini
• Priming sites
(From Molecular cloning 3, 8.14)

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Primer design

1. 1. PCR primer design • Local software Primer premier5, Oligo 6.1 • Web-based design Primer3 (Whitehead Institute/MIT Center for Genome Research)

2. Primer premier5

3.

4.

5.

6.

7.

8.

9.

10.

11. Principles for primer design • Base composition • Length • Repeated and self-complementary seq. • Cross dimer • Tm • 3’ Termini • Priming sites (From Molecular cloning 3, 8.14)