This service is the fastest way for you to obtain sequence information from a plasmid clone, while saving the time, effort and expense of performing plasmid preps. You can go from colony to sequence in as little as 20 hours!
By requesting our rush RCA24 Service, you can go from colony to sequence in 20 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.
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.
Amplified templates can be directly sequenced without further purification – saving you time and money*.
|Single Colonies on Agar Plates (Sequetech Picks the Colonies)||After analyzing the sequences, mini-cultures of the clones of interest can be returned (by request.)|
|Single Colonies from Agar Plates (Customer Submits Picked Colonies)||Cells should be picked into 10ul growth media supplemented with the appropriate antibiotic (please identify the antibiotic.) Make sure all tubes and plates are tightly sealed and do not leak. Please do not freeze.|
|Liquid Cultures and Glycerol Stocks||We request 10ul of culture grown with the appropriate antibiotic (please identify the antibiotic.) Make sure all tubes and plates are tightly sealed and do not leak. Please do not freeze.|
|Partially Purified or Dilute Plasmid Preps||Please supply 10ul of DNA at concentrations as low as 10pg/ul.|
|4 pmole/primer extension||2 uM||Submitting extra will allow for optimization and troubleshooting. Primers and templates are stored for at least 3 months.|