What are the key features of a TaqMan™ MGB probe?

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Answer

TaqMan™ MGB probes have a nonfluorescent quencher at the 3´ end and a minor groove binder at the 3´ end that increases the melting temperature (Tm) of probe, allowing the use of shorter probes.  For more information on MGB probes, please consult the following manual: "Primer Express v1.5 and TaqMan™ MGB Probes for Allelic Discrimination: All PCR Instruments: User Bulletin." You can find a copy on our website by entering this title in the main search box.

Answer Id: E1371

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When using the TaqMan™ Gene Signature Plates, should I mix the reactions after adding the master mix and template to the wells containing lyophilized primers and probe?

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Answer

No, but it is recommended that the plate be centrifuged to draw down the liquid to the bottom of the wells.

Answer Id: E1370

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Can TaqMan™ Gene Expression Assays be used in a multiplex reaction?

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Answer

These assays have not been tested to run in a multiplex environment. If you would like to multiplex TaqMan™ Gene Expression Assays, you can select any of our pre-design assays with a VIC dye label so that you can run two seperate assays in the same well. For information about optimizing a multiplex gene expression experiment, please see "Factors Influencing Multiplex Real-Time PCR". You can get the document by using "Factors influencing multiplex' as a keyword to search our web site.

Answer Id: E1836

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What are your recommendations for the best way to dilute and store my TaqMan™ probe?

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Answer

The probe should be diluted in TE buffer [10 mM Tris-HCl, pH 8.0 (at 25°C), 1 mM EDTA]. We recommend storing your probes, both stocks and working solutions, at -20°C in aliquots. This is to cut down on the number of times the probe is subject to freeze-thaws. For detailed information on diluting primers and probes, please refer to the tutorial entitled "Reconstituting and Diluting Primers and TaqMan™ Probes". You can find it on our website by entering this title in the main search field.

Answer Id: E1129

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