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?
What are the key features of a TaqMan® MGB probe?
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: 1371
Can TaqMan® Gene Expression Assays be used in a multiplex reaction?
These assays have not been tested to run in a multiplex environment. In addition, each assay is pre-formulated with a 6-FAM™ reporter dye, so it is not possible to incorporate two different TaqMan® Gene Expression Assays into a multiplex reaction. If you would like to multiplex TaqMan® Gene Expression Assays with an endogenous control gene, our TaqMan® Endogenous Control product line offers some commonly used housekeeping genes for human, mouse, rat, and other species, and one assay that will detect 18S rRNA from all eukaryotic organisms. These assays are available with a VIC® reporter dye and in primer-limited formulations suitable for multiplexing. To see the list of these assays, please type "TaqMan Endogenous Control" in the web search form to search and it will lead you to the product page. For information about optimizing a multiplex gene expression experiment, please see "User Bulletin #5: Multiplex PCR with TaqMan® VIC® Probes (P/N 4306236B)". You can get the document by using "User Bulletin #5' as a keyword to search our web site. This User Bulletin was written for use on the ABI PRISM® 7700, but the concepts are valid for all of our real-time PCR instruments.
Answer Id: 1836