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?
-
-
Hi Creeves
I wondering why you are using RCA with Nextera. Nextera XT have been designed for library preparation directly from plasmids among other types of DNA. I see RCA a shortcut to avoid plasmid preps. -
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.Comment
-
NGS has revolutionized clinical microbiology by offering new ways to identify and surveil pathogens without prior knowledge of the target. One popular approach is 16S rRNA and ITS amplicon sequencing, which targets specific microbial markers. But studies are increasingly incorporating shotgun metagenomics, a technique that sequences all DNA from a sample without targeting specific microbial markers. This approach allows more detailed taxonomic classification, detection...Yesterday, 06:46 AM -
Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?
Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating...10-14-2026, 09:55 AM
Comment