TaqMan™ miRNA ABC Purification Kit - Human Panel B - FAQs

I am using the TaqMan miRNA ABC Purification Kit and am having difficulty detecting miRNA in qPCR assays. What is the issue?

Here are possible causes and solutions:

- RNA degradation due to RNAse contamination during procedure: See “Guidelines to prevent RNAse contamination” on page 12 of the manual (https://tools.thermofisher.com/content/sfs/manuals/4473439C.pdf) for proper handling of sample.
- Inefficient elution of miRNA from the beads: follow hybridization, wash, and elution steps on pages 17-18.
- Inefficient washing: fully resuspend beads in wash buffers, and completely remove buffers at the end of the washes.
- Beads dried out during procedure: prepare the beads just before use and keep the beads suspended in liquid and the tube cap closed in between steps.
- Buffers not diluted as directed: prepare buffers as directed on page 13.
- Kit components compromised due to inappropriate storage conditions: store reagents as directed in “Kit contents and storage conditions” on page 9 of the manual.