Flexprep: scale-flexible rapid plasmid preparation for analysis of recombinant clones.
A scale-flexible and cost-efective protocol for plasmid preparation is described to cover miniprep and midiprep scale work in a microcentriguge format for analysis of recombinant clones, this protocol relies on a modified alkaline lysis of Escherichia coli cells and subsequent purification of plasmid DNA with no organic extraction and alcohol precipitation. It can process up to 20 mL of E. coli cells carrying 3-10 kbp plasmid vectors in < 10 min. Flexprep delivers sufficient yield and purity of plasmid DNA for routine applications including restriction enzyme digestion and fluorescent automated sequencing.
Aranishi
Futoshi
F
Molecular Biology Division, National Institute of Fisheries Science, Kanazawa, Yokohama, Japan. aranishi@affrc.go.jp
D004247:DNA / Q000302:isolation & purification; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D010957:Plasmids / Q000302:isolation & purification*
D004247:DNA