Millipore Technical Publications |
 | UG-P36315 | |
Montage Plasmid MiniprepHTS Kit User Guide | ||
| Lit No.: | P36315 | |
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| The Montage Plasmid MiniprepHTS Kit provides all of the reagents and materials necessary to purify plasmid or bacterial artificial chromosome (BAC) DNA using a simple protocol that eliminates lengthy bind/elute methods and centrifugation. Employing unique separation technology, Millipore has developed a line of easy-to-use DNA miniprep kits that yield DNA suitable for the most sensitive downstream applications. In addition, this technology has significantly reduced the time required for processing samples. Following bacterial lysis, three short filtration steps are all that is required to prepare 96 clean DNA samples from each plate. The DNA is retained by Millipore’s proprietary size-exclusion membrane while proteins and contaminants are filtered through to waste. Current genome sequencing strategies require the cloning of large fragments of genomic DNA into F factor-based BAC vectors. One obstacle when adapting this strategy to a high throughput format is the low copy number of these vectors. Because there are only 1–2 copies of each BAC maintained per cell, the theoretical yield of BAC DNA from a given volume of culture is significantly lower than that for plasmid DNA. In addition, the large variability in insert size contributes proportionally to the variability in BAC DNA yield. Following the recommendations provided in this protocol to optimize bacterial cell culturing conditions will result in a sufficient yield of high quality DNA for desired downstream applications. | |
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