What reagents and/or kits are required for the TaqMan® Advanced miRNA Assay workflow using the TaqMan® Advanced miRNA Human A and B cards (Cat. No. A31805)?
All materials required for TaqMan® Advanced miRNA Assay workflow using TaqMan® Advanced miRNA Human and B cards (Cat. A31805) can be found on pages 9-11, in the TaqMan® Advanced miRNA Assays User Guide.
Reagents and kits required for the TaqMan® Advanced miRNA Assay workflow include, but are not limited to:
• RNA Isolation Kit (see recommended kits on page 9) • TaqMan® Advanced miRNA cDNA Synthesis Kit (Cat. No. A28007) • TaqMan® Fast Advanced Master Mix (Cat. No. 4444557)
Please refer to the user guide linked above for a full list of other materials and equipment required for the workflow.
I am not seeing good amplification with my TaqMan Advanced miRNA array cards. What is wrong?
The TaqMan Advanced miRNA array cards allow one to profile over 300 different miRNA targets on one card. Due to the large number of targets and the nature of your sample, it is possible that some miRNA on the card will not be expressed on your sample, or expressed at very low levels. This is one reason why you will not see good amplification plots for every spot on the array card. Another possibility is the sample quality or input. Make sure to check the concentration and quality of the RNA sample before reverse transcription. Check the Ct values for your endogenous control assays on the card, and see if it is in the expected range for your typical samples.