The Rapid Plasmid Midiprep Kit combines the power of HiBind® technology with the time tested consistency of alkaline-SDS lysis of bacterial cells to deliver high quality plasmid DNA. The midi-columns facilitate the binding, washing, and elution steps thus enabling multiple samples to be simultaneously processed. HiBind® matrix that avidly, but reversibly, binds DNA under specific conditions allowing proteins and other contaminants to be removed. Nucleic acids are easily eluted with de-ionized water or low salt buffer.

Procedure

Pellet cells from 50 - 100 ml of overnight culture, depending on cell density. Re-suspend collected cells in Buffer I. Add Solution II and RNase to effectively lyse bacterial cells. Addition of Solution III neutralizes the reaction, enabling the DNA to bind to the HiBind® column . Pellet chromosomal DNA and cellular debris, by a brief centrifugation step, and pass the cleared lysate through the column to bind plasmid DNA. Wash Buffers HB and DNA wash to remove impurities. Dry the column with an additional centrifugation step and elute plasmid DNA with sterile de-ionized water or TE buffer for further use.

Applications

Rapid Plasmid Midi Kit provides high quality, reproducible plasmid DNA for use in the following applications :

• Automated fluorescent DNA sequencing
• Radioactive sequencing
• Restriction digestion
• Screening
• Ligation and transformation

Sequencing autoradiogram of plasmid DNA isolated by using Rapid Plasmid Midi Kit