Archives of Histology and Cytology
Original articles
Atomic force microscopy analysis of rolling circle amplification of plasmid DNA
Ryushin MizutaMidori MizutaDaisuke Kitamura
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Volume 66 (2003) Issue 2 Pages 175-181

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Rolling circle amplification (RCA) of plasmid DNA using random hexamers and bacteriophage phi29 DNA polymerase is an increasingly applied technique for amplifying template DNA for DNA sequencing. We analyzed this RCA reaction at a single-molecular level by atomic force microscopy (AFM) and found that multibranched amplified products containing tandem repeats of a circle unit are formed within 1 h. We also used the RCA product of a GFP expression vector for the protein expression in cells, and found that the crude RCA product from one bacterial colony is sufficient for the GFP expression. Thus, the RCA reaction is useful in amplifying DNA for both DNA sequencing and protein expression.

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© 2003 by International Society of Histology and Cytology
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