GLOBINclear™-Human Kit, for globin mRNA depletion - FAQs

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2 个常见问题解答

I used the GLOBINclear Kit to deplete globin mRNA from my total RNA sample, but the depletion was not very efficient. What should I do?

To maximize removal of globin mRNA, follow the procedure exactly. Also pay attention to the most crucial parts of the procedure listed below:

-The GLOBINclear Kit procedure is optimized for depletion of globin mRNA from 10 µg or less total RNA from human, mouse, or rat whole blood. Using more than 10 µg RNA in the procedure will compromise removal of globin mRNA.
-Be sure to follow the temperature and time recommendations for the hybridizations in steps II.D.4 on page 9 and II.E.2 on page 10 of the manual (https://tools.thermofisher.com/content/sfs/manuals/fm_1980.pdf) for optimal depletion of globin mRNA.

Does the GLOBINclear Kit deplete ribosomal RNA (rRNA) in addition to globin mRNA?

The GLOBINclear Kit does not contain capture probes for ribosomal RNA (rRNA) sequences; however, after globin mRNA depletion with the kit, the percentage of ribosomal reads for the kit is only 1% of total reads (https://pubmed.ncbi.nlm.nih.gov/33308163/). This low percentage is irrelevant for most downstream applications.