Can I check the end product of a TaqMan Gene Expression Assay by running an agarose gel?
Yes, you can check the end product on an agarose gel. If you use the TaqMan Universal PCR Master Mix (including AmpErase UNG, Cat. No. 4304437), we recommend running the gel immediately after the PCR, since the trace amount of AmpErase UNG may digest the end product that contains double-stranded DNA with dUTP incorporated. The average length of amplicon ranges from 50-150 bp. The exact amplicon length is indicated on the detailed page of each assay on our website. You can get to that page by clicking on the Assay ID; the amplicon information is listed in the section "Assay Details".
How do I select a TaqMan Gene Expression Assay for an endogenous control in relative quantitative PCR (qPCR)?
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
How can I reorder a Custom TaqMan Assay that I ordered previously?
To reorder the same Custom TaqMan Assay, you can follow these steps:
- Select an appropriate catalog number, considering the size of the assay that you want. You can find the different catalog numbers for Custom TaqMan Assays on their respective product pages on thermofisher.com.
- Once you have identified the necessary catalog number, go to www.thermofisher.com and click on "Quick Order", in the top right corner of the screen.
- In the following page, paste your selected catalog number in the "Catalog Number" field and paste the assay ID from your previous order in the "Assay ID" field.
- Click "Add to cart" and then in the following screen click "Proceed to check out".
Can I perform a one-step RT-PCR reaction using TaqMan Gene Expression Assays?
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.
When preparing the TaqMan or Custom TaqMan Gene Expression Assays, what is the recommended reaction volume?
When preparing the TaqMan or Custom TaqMan Gene Expression Assays, the recommended and supported volume is 20-50 µL for a 96-well standard plate set-up; 10-20 µL for a 96-well fast plate set-up and 5-10 µL for a 384-well set-up. For more information, please consult the TaqMan Gene Expression Assays and Custom TaqMan Gene Expression Assays protocols.
Find additional tips, troubleshooting help, and resources within our TaqMan Primers and Probes Support Center.
