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View additional product information for Custom TaqMan™ Gene Expression Assay, FAM - FAQs (4332078, 4332079, 4331348)
23 product FAQs found
An assay or sample may be deleted from a study only if there is no data or wells associated with it. Upon import of an experiment, the software collects all the assays and samples from the plate and lists them in the Setup > Assays or Setup > Samples workspaces. The assays and samples are stored in these workspaces as a library, and remain there even if you delete the experiment from the Study. Deleting the experiment will remove any data (wells) associated with the assays or samples, but not the assays or samples from the library. The assays and samples must then be deleted from these workspaces to remove them from the Study.
For optimal results, we recommend to run the reaction plate right after setting up the reaction. If a reaction plate cannot be run within 2 hours of completing the reaction setup, then freeze/refrigerate the reaction plate until it can be loaded and run on the fast PCR instrument platform.
No, we do not recommend this approach. Since the standard TaqMan Universal PCR Master Mix is not optimized for running with fast thermal cycling, assay results will be significantly compromised.
The TaqMan Fast Universal PCR Master Mix is provided without any UNG and you should add in AmpEraseUNG to reactions separately, with the addition of an initial 2 minute cycling step at 50º C. If you prefer to have a master mix with the UNG pre-added, please use TaqMan Fast Advanced Master Mix.
In general, assays have been found to perform comparably with these reagents, but they may not provide exactly identical results.
We recommend a storage temperature of 4C and the shelf life is 9 months from the manufacturing date.
We recommend adding 10 to 100 ng of RNA converted to cDNA as your template.
No. The only Fast reagents that we offer are the TaqMan Fast Universal PCR Master Mix (2X) for single plex use, and Fast Advanced Master Mix for greater sensitivity and multiplexing use.
TaqMan Fast Universal PCR Master Mix is recommended for two-step RT-PCR, but for one-step reactions, we recommend using TaqMan Fast Virus 1-Step Master Mix as the preferred reagent.
The hot-start DNA polymerase enzyme system in the TaqMan Fast Universal PCR Master Mix does not require an activation step, and can be activated with significantly shorter hold times for the thermal cycling stages of PCR, thereby minimizing PCR run time. For more information on using this reagent, please consult the TaqMan Fast Universal PCR Master Mix (2X) protocol.
The fast master mix has demonstrated detection down to 10 copies of the RNase P gene with genomic DNA. Of course, detection limits may vary depending on factors such as assay design and sample preparation.
The following research assays are supported using the TaqMan Fast Universal and/or Advanced Fast PCR Master Mix: TaqMan Gene Expression Assays, Custom TaqMan Gene Expression Assays, Custom primers and probes designed from Primer Express software-designed quantitative assays.
The primer and probe sequences for your custom TaqMan Gene Expression Assay can be found in the Assay Information File (AIF) which can be downloaded here. Once there, select the option "TaqMan™ or Absolute Q™ Assay" and, on the next page, enter the Sales Order number and the Rack/Plate ID for the respective assay (these numbers can be found on the assay packaging near the barcode), and submit. From there you can download a folder including a txt-file with all sequences.
Please note that the sequences are only accessible once the assays have been received.
Find additional tips, troubleshooting help, and resources within our TaqMan Primers and Probes Support Center.
Please see the TaqMan Gene Expression Assays - single-tube assays User Guide , page 8.
An endogenous control gene is expressed at a consistent level in all studied conditions. It is impossible to predict exactly how any gene will behave under a given range of conditions. The best way of selecting the most appropriate control gene for a relative qPCR experiment is to select some candidate genes and determine their expression levels across the range of experimental conditions and treatments. The genes most stably expressed across these conditions will be the most appropriate controls. See the following links on selection and usage.
Check the TaqMan Endogenous Control Assay Selector Tool here.
Find additional tips, troubleshooting help, and resources within our Gene Expression Support Center
While TaqMan Gene Expression Assays can be used to perform a one-step RT-PCR reaction, these assays have not been optimized for this application. TaqMan Gene Expression Assays have been designed, tested, and optimized using Applied Biosystems universal conditions for a two-step RT-PCR reaction.
The details for custom TaqMan assays can be downloaded at: https://www.thermofisher.com/order/taqman-files. Select the option "TaqMan Assays" and, in the screen that follows, indicate the sales order number and the Rack ID of your assay.
Both these numbers can be found on the assay package, next to the barcode.
The sequence provided for the assay design may contain mismatches with sample sequences. We recommend that you perform bioinformatics analysis for assay design.
For more information, see the Custom TaqMan Assays Design and Ordering Guide (Pub.No. 4367671).
For orders placed after March 2016, assay information files can be accessed from this page: www.thermofisher.com/taqmanfiles
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
The number of reactions will depend on your final reaction volume. The Custom and Custom Plus TaqMan RNA assays are supplied with the following volumes:
Small: 360 µL (20X)
Medium: 750 µL (20X)
Large: 976 µL (60X)
Assuming a 20 µL final reaction volume, the small size would provide 360 reactions, the medium size would provide 750 reactions, and the large size would provide 2,900 reactions.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
The Custom and Custom Plus TaqMan RNA assays are supplied with the following volumes:
Small: 360 µL
Medium: 750 µL
Large: 976 µL
Please note that the large scale is supplied in a 60X format, while the small and medium scales are supplied in 20X format.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Custom and Custom Plus TaqMan RNA assays can be ordered online with either FAM, VIC, or VIC primer limited.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Custom and Custom Plus TaqMan RNA assays can be ordered online with either FAM or VIC dye, with a MGB-NFQ quencher.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.