Supplementary data: In vitro amplification of whole large plasmids via transposon-mediated oriC insertion

<b>Supplementary
Table: A list of primers used in this study</b>
<b>Supplementary
Figure 1. Amplification of a small plasmid contaminant in isothermal RCR.</b> The
indicated amount of pRpoABCDZ was subjected into the Tn-oriC insertion
reaction, followed by RCR at 30˚C for 16 h.
<b>Supplementary
Figure 2. Restriction enzyme analysis of the amplification products of pTT8.</b> Purified pTT8 plasmid or the amplification
product of the pTT8 plasmid generated by Tn-RCR were incubated at 37ºC for 3
h with (cut) or without (no cut) <i>Kpn</i>I and <i>Nhe</i>I. The incubated
products were then analyzed by 1% agarose-gel electrophoresis and SYBR Green
staining. It should be noted that the ratio of supercoiled form decreased in
“no cut” sample due to DNA damage during the incubation. Size-marker
fragments (M2) were derived from lambda phage DNA. A digestion map of pTT8 is
shown on the right.

<b>Supplementary Figure
3. Restriction enzyme analysis of the amplification products of the F plasmid.</b> A purified F-plasmid
or the amplification product of the F plasmid generated by Tn-RCR were digested
with <i>Pme</i>I as described in Supplementary Figure 2. The digested products
were analyzed by 0.75% pulse-field agarose gel electrophoresis using a Pippin
pulse power supply (Sage science) and SYBR Green staining. Although this
pulse-field methods can separate large sized linear DNA, discrimination of
supercoiled DNA band is difficult. Size-marker fragments (M3), <i>Saccharomyces
cerevisiae</i> chromosomal DNA. Size-marker fragments (M4) were derived from T7
phage DNA. A digestion map of F plasmid is shown on the right.