-Speeding Up-

@plasmid extraction

Centrifugation (10000 rpm, 1 min, 4℃), and disposed supernatant.

Add 100μl solution I and vortex.

Add 200μl solution II and upside down 5 times.

Add 150μl solution III and upside down 5 times, put on ice for 5 min.

Centrifugation (15000 rpm, 5 min, 4℃), and disposed supernatant.

Add 95% EtOH twofold volume and leave room temperature for 2 min.

Centrifugation (15000 rpm, 5 min, 4℃), and disposed supernatant.

Cleaned by 70% EtOH

Centrifugation (15000 rpm, 2 min, 4℃), and disposed supernatant, and drying.

Add TE

@restriction enzyme

Add Xho I 1μl, M buffer 3μl, SDW 16μl, plasmid solution 10μl to tubes, and incubated at 37℃ for 1 hour

@electrophoresis

Add gel to electrophoretic apparatus, maker and suspended by 1μl Loading buffer, and electrophored in 100V, 30min.

Then confirmated bands in UV and took a photo.

move E.coli to kanamycin culture media.