Rapid extraction of plasmid pGT5 from the hyperthermophilic archaeon Pyrococcus abyssi.
Hyperthermophilic archaea, specifically Pyrococcus spp., are the target of current efforts in developing heterologous expression systems. However, the published plasmid purification and plasmid screening protocols are long and tedious. We describe a fast, simple protocol for plasmid purification from Pyrococcus spp. developed while extracting the plasmid pGT5 from Pyrococcus abyssi cells. The protocol is modified from the procedures for commercial plasmid minipreps and is completed in about 20 min. The DNA is easily digested by restriction enzymes and can be used in sequencing reactions without additional purification.
González
J M
JM
Center of Marine Biotechnology, University of Maryland, Biotechnology Institute, Baltimore 21202, USA.
Robb
F T
FT
D004262:DNA Restriction Enzymes / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D010957:Plasmids; D019711:Pyrococcus / Q000235:genetics*
D004262:DNA Restriction Enzymes