High-speed plasmid isolation using 96-well, size-exclusion filter plates
Harris, D.; Engelstein, M.; Parry, R.; Smith, J.; Mabuchi, M.; Millipore, J.Leonard.
Biotechniques 32(3): 626-8 630-1
2002
ISSN/ISBN: 0736-6205 PMID: 11911665 Document Number: 551322
The accelerating pace of genomics analysis has necessitated the abbreviation of DNA sample preparation protocols. We have developed a size-exclusion-based system for the rapid isolation of plasmid DNA in a 96-well microplate format. This high-speed protocol employs a modified alkaline lysis method for the preparation of the bacterial lysate, followed by three short vacuum filtration steps. Unlike traditional bind/wash/elute methods, there is no need to use chaotropic salts or ethanol. The samples are recovered from the top side of the MultiScreen96(R) PLASMID plates. Starting with bacterial cell pellets, the entire procedure for purifying the plasmid DNA can be performed in 30 min with a multichannel pipettor. The high yields, reproducibility, and quality of the plasmids make this system a good choice for any cloning or DNA sequencing operation.