QIAGEN Plasmid Kits

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Overnight cultures (100 ml) of B. xylanisolvens DSM 23964, Escherichia coli DSM 3876 (plasmid RP4, 60 kb), and E. coli DSM 6202 (plasmid pSC 101, 9.4 kb) were harvested by centrifugation for 10 min at 13,000 × g at 4°C and washed three times in 2 ml sodium phosphate buffer (50 mM, pH 7.0).	Plasmid DNA was extracted with the Qiagen plasmid midikit (Qiagen) according to the manufacturer's protocol. According to the Qiagen manufacturer, the plasmid purification kit used in this study was able to purify plasmids up to approximately 150 kb.

Upstream tips
Overnight cultures (100 ml) of B. xylanisolvens DSM 23964, Escherichia coli DSM 3876 (plasmid RP4, 60 kb), and E. coli DSM 6202 (plasmid pSC 101, 9.4 kb) were harvested by centrifugation for 10 min at 13,000 × g at 4°C and washed three times in 2 ml sodium phosphate buffer (50 mM, pH 7.0).
Protocol tips
Plasmid DNA was extracted with the Qiagen plasmid midikit (Qiagen) according to the manufacturer's protocol. According to the Qiagen manufacturer, the plasmid purification kit used in this study was able to purify plasmids up to approximately 150 kb.

Publication protocol

Overnight cultures (100 ml) of B. xylanisolvens DSM 23964, Escherichia coli DSM 3876 (plasmid RP4, 60 kb), and E. coli DSM 6202 (plasmid pSC 101, 9.4 kb) were harvested by centrifugation for 10 min at 13,000 × g at 4°C and washed three times in 2 ml sodium phosphate buffer (50 mM, pH 7.0). Plasmid DNA was extracted with the Qiagen plasmid midikit (Qiagen) according to the manufacturer's protocol. According to the Qiagen manufacturer, the plasmid purification kit used in this study was able to purify plasmids up to approximately 150 kb. The analysis of plasmid DNA was performed by a modification of the procedure as previously described (4). The quantitative analysis of plasmid DNA was done by horizontal agarose gel electrophoresis in 1% agarose gel.

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Manufacturer protocol

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