
DRAFT
December 16, 2004 3:54 pm, GSG_Chapter_4-02.fm
Chapter 2 Designing Primers and Probes for Quantification Assays
Manually Designing Primers and Probes
Primer Express Software 3.0 Getting Started Guide 13
Notes
2
Manually Designing Primers and Probes
You may choose to manually design primers and probes for various reasons:
• Automated primer/probe design did not find primers and probes.
• To design a probe over an exon junction.
• To design a probe for DNA sequence homologs.
• To design primers and probes according to your own specifications.
Creating a
Quantification
Document
Create a Quantification document and load a sequence file as you would for automatic
primer/probe set design. See “Creating a Quantification Document” on page 9.
Manually
Designing the
Probe
1. Select a putative probe region containing at least 25 bases.
2. Copy (Ctrl+C) the sequence.
IMPORTANT! The Primer Probe Test Tool eliminates non-ATCG bases. Before copying
a sequence, change any non-ATCG bases, or select a different region of the sequence.
3. Select Too ls > P rime r Probe Te st Too l. The Primer Probe Test Tool dialog box
appears (see “Figure 4. Primer Probe Test Tool dialog box” on page 14).
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