实验概要

The PureLink™ HiPure Plasmid DNA Midiprep Kit allows purification of 100–350 μg of high-quality plasmid DNA from 15–25 mL overnight E. coli cultures in ~2 hours when cloning high copy number plasmids.

主要 试剂 Before Starting Before beginning, verify that the Resuspension Buffer (R3) contains RNase A, and no precipitate has formed in the Lysis Buffer (L7). Materials Needed Overnight culture of transformed E. coli cells Isopropanol 70% ethanol Sterile, microcentrifuge tubes PureLink™ Nucleic Acid Purification Rack Tubes or centrifuge bottles for harvesting cells Centrifuge and rotor appropriate for harvesting cells Appropriate 15-mL centrifuge tubes capable of withstanding centrifugation forces >12,000 × g Centrifuge capable of centrifuging at >12,000 × g at 47°C Optional : PureLink™ HiPure Precipitator Module Components Supplied with the Kit Resuspension Buffer (R3) with RNase A Lysis Buffer (L7) Precipitation Buffer (N3) Equilibration Buffer (EQ1) Wash Buffer (W8) Elution Buffer (E4) TE Buffer (TE) HiPure Filter Midi Columns Column Holder Note: The protocol for the Midiprep kit of the PureLink™ HiPure Plasmid Filter Purification Kits is designed for purification of both high and low copy number plasmids without the need for adjusting buffer volumes.