title: A high-salinity solution with calcium chloride enables RNase-free, easy plasmid isolation within 55 minutes.
abstract: We dramatically improved a plasmid-isolation protocol based on the popular alkaline-sodium dodecyl sulfate plasmid isolation method. Our modified method provides significant time and cost savings. We used a modified solution during the neutralization step, which allowed us to skip several subsequent handling steps, saving a great amount of time. The plasmids purified by this method were of high quality, and the optical density ratio 260 and 280 was approximately 1.8. Plasmid DNA isolated by our method was of sufficient quality to perform subsequent restriction enzyme cuts and other downstream experiments, including budding yeast transformation, cultured cell transfection, and Caenorhabditis elegans injection experiments.
authors.affiliation: Department of Applied Biochemistry, School of Engineering, Tokai University, Kanagawa, Japan.
authors.forename: Noboru
authors.initials: N
authors.lastname: Sasagawa
authors.forename: Michinori
authors.initials: M
authors.lastname: Koebis
authors.forename: Yoji
authors.initials: Y
authors.lastname: Yonemura
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Mitsuhashi
authors.forename: Shoichi
authors.initials: S
authors.lastname: Ishiura
mesh_terms: D002122:Calcium Chloride / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry* / Q000302:isolation & purification*; D012260:Ribonucleases / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry*
chemical_list: D012965:Sodium Chloride; D012260:Ribonucleases; D002122:Calcium Chloride