Note that RCA adds an additional 24 hours to the sequencing turnaround time. RCA is recommended for circular templates less than 15 kb in length.
Dean, Frank B. et al. “Rapid Amplification of Plasmid and Phage DNA Using Phi29 DNA Polymerase and Multiply-Primed Rolling Circle Amplification.” Genome Research 11.6 (2001): 1095–1099.
*Please note: RCA protocol is intended for circular templates <10 kb; linear templates will not be amplified sufficiently for direct Sanger sequencing.
Longer circular templates, or circular templates carried in bacterial strains with functional endA may not be reliably amplified, contributing to variable results
Please visit our Sample Submission Guidelines for bacteria and phage templates for additional details.