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 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.
How much cDNA is recommended per TaqMan or Custom TaqMan Gene Expression reaction?
For optimal performance of the TaqMan and Custom TaqMan Gene Expression Assays, use 1 to 100 ng of cDNA per 50 µL total reaction volume. For more information, please consult the TaqMan Gene Expression Assay protocol and Custom TaqMan Gene Expression Assay protocol.
Find additional tips, troubleshooting help, and resources within our TaqMan Primers and Probes Support Center.