RCA Service

Sequetech DNA SequencingThis service is the fastest way for you to obtain sequence information from a plasmid clone, while saving the considerable time, effort and expense of performing plasmid preps.   We offer two levels of service, RCA24 and RCA48.  By requesting our rush RCA24 Service, you can go from colony to sequence in 18 to 24 hours!

RCA is an acronym for Rolling Circle Amplification and is an in vitro process whereby circular DNA from a variety of sources can be amplified to produce high quality sequencing templates.  Suitable input DNA includes partially purified or dilute DNA preps, liquid bacterial cultures or bacterial colonies.  Amplified templates can be directly sequenced without further purification.

Each RCA reaction yields sufficient template to perform up to 3 sequencing reactions with no additional RCA charges. (Sequencing fees charged separately.) For sequencing requests involving more than 3 single primer extensions from an RCA'd template, additional RCA reactions must be performed to obtain enough template and therefore will cost more.

RCA has been widely utilized to amplify DNA for a variety of purposes, including sequencing template preparation.

In RCA, DNA synthesis from a circular DNA (primed by random hexamers and catalyzed by Phi29 DNA polymerase) produces single-strand, linear concatenated copies of the circular sequence.  This primary product subsequently acts as template for further DNA synthesis with the products, in turn, also serving as templates for further synthesis.  The combined result of this process is exponential amplification of the original circular DNA.  Please see Figure 1.

Figure 1.  RCA Replicating DNA 

Sequetech will, using RCA, amplify and sequence your templates.  If requested, we will also pick colonies to serve as a source of your clones.

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