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Assessment of Long IVT mRNA Fragments with the Agilent Fragment Analyzer System
Poster Title: Assessment of Long IVT mRNA Fragments with the Agilent Fragment Analyzer System
Submitted on 16 Dec 2020
Author(s): Chava Pocernich, Kyle Luttgeharm, and Steve Siembieda
Affiliations: Agilent Technologies
This poster was presented at AMP 2019
Poster Views: 584
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Poster Information
Abstract: Through the process of in vitro transcription (IVT), high quality long sequence RNAs can be synthetically produced at high concentrations. IVT mRNA is becoming widespread in research areas such as ribozyme and aptamer synthesis, mRNA synthesis, RNA interference, and antisense RNA techniques. Conventionally, synthesized RNA fragments are less than 100 nt in length, but research in gene structure and functional studies requires extremely long RNA. To date, there are no pre-made kits for sizing RNA over 6,000 nt. Thus, to analyze long IVT RNAs, an extended separation method was developed for the Agilent 5200 Fragment Analyzer using a commercially available RNA ladder. The method provides enhanced resolution aiding in the detection of minor degradation, contamination, or sizing differences from incomplete transcription. Using this method, 9,000 and 10,000 nt IVT mRNA samples were successfully analyzed with good sizing precision and accuracy, demonstrating that a commercially available RNA ladder can be used with the Fragment Analyzer system to provide accurate sizing of large IVT mRNA samples.

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