AMX DESIGN XPRESS V 1.5 - PROGRAMMER GUIDE Manuel d'utilisateur télécharger pdf (Page 30)

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December 16, 2004 3:54 pm, GSG_Chapter_4-02.fm

Chapter 2 Designing Primers and Probes for Quantification Assays

Manually Designing Primers for SYBR

Green Dye Assays

Primer Express Software 3.0 Getting Started Guide 23

Notes

Manually Designing Primers for SYBR

Green Dye Assays

You may choose to manually design primers and probes for a various reasons:

• Automated primer/probe design did not find primers.

• To design primers 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

Primers

To design the Forward Primer:

1. In the Sequence tab, select a putative forward primer sequence 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 7. Primer Probe Test Tool dialog box” on page 24).

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AMX DESIGN XPRESS V 1.5 - PROGRAMMER GUIDE Manuel d'utilisateur Page 30