We were thinking of using rolling circle amplification of plasmids to generate the DNA we would use to make Nextera libraries for high plexity pools to load on a MiSeq. We have succeeded in sequencing 384 plasmids in one run and are moving to 1536 and beyond. Has anyone out there tried RCA with the Nextera process?
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The reason is simply cost reduction. We want base pair resolution QC of the thousands of constructs we routinely make in our automated strain engineering pipeline. The MiSeq can reliably sequence about 2000 10-20 kb plasmids in one run, but at that scale the plasmid prep cost becomes an issue.
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The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist...-
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