This technical note highlights the challenges faced in quality control for RNA therapeutics, an emerging area of gene therapy. Traditional methods, such as RT-PCR and RT-qPCR, have limitations when detecting poly(A) tail length and can introduce artifacts, preventing accurate and precise quantitation. An ideal quality control approach should allow for a comprehensive evaluation of full-length products, intermediates, and byproducts.
In this tech note, Azenta Life Sciences presents a novel approach to sequencing full-length, manufactured RNA molecules using PacBio long-read technology. This method preserves the entire length of the poly(A) tail, enabling sensitive and specific counting of the poly(A) tail length. The results demonstrate that this next-generation sequencing approach offers read depth sufficient to identify and confirm overall sample purity, including intermediate and alternative products present within the sample, and ensures complete fidelity of the manufactured RNA therapeutic.
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