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In the past 20 years of research, circular RNA (circRNA) has been found to exist in all types of living organisms in different types. This round ribonucleotide sequence is much richer than previously thought, but we know very little about the exact function of circRNA. Researchers use complex sequencing techniques and bioinformatics analysis to identify and discover circRNA and hope to construct related models by in vitro preparation of circRNA for more in-depth functional studies.
IntegrateRNA, a leading biotechnology company with years of experience in RNA research, utilizes the latest T4 ligase synthesis system to provide customized circRNA in vitro synthesis services for basic research, disease pathology research, and drug discovery research. You can submit a target circRNA sequence or use our design tool to configure the circRNA sequence. We are committed to providing our customers with the most reliable and effective research services.
➢ Strategy design
➢ In vitro transcription and product purification
➢ Dephosphorylation of RNA and 5′-Phosphorylation
➢ Substrates with 5'-monophosphate and free 3'-hydroxyl are required due to enzymatic ligation of RNA. Therefore, we dephosphorylate the 5'-0H group transcribed in vitro to a monophosphate by polynucleotide kinase and ATP.
➢ As an alternative, we can use GMP instead of GTP as the first nucleotide to elicit a new RNA that contains GMP and can be used directly for ligation.
➢ RNA circularization — T4 RNA ligase
As an alternative to chemical RNA synthesis, enzymatic protocols can produce long RNAs that may be in the kb range.
➢ Intermolecular end-linking is a serious side reaction and cannot be completely inhibited. The cyclic product can be isolated from any oligomer species by polyacrylamide gel electrophoresis.
IntegrateRNA, a global leading biotechnology company, is dedicated to providing products and services to a wide range of genomic researchers, including various types of RNA discovery, analysis, and related bioinformatics services. We are committed to providing our customers with the most reliable and efficient research services to best meet your research goals, with reduced turnaround time and lower prices. If you have any questions, please feel free to contact us.