Millipore Technical Publications
Plasmid Minipreparation using MultiScreen 96-Well Filter Plates
|The need for rapid, cost effective high throughput methods for plasmid preparation has been steadily increasing. This technical note details a protocol enabling the use of MultiScreen filtration plates and user-supplied reagents to complete plasmid purification in the 96-well format without the need for expensive kits. The resultant purified plasmid DNA is of high quality and fully functional as assayed by cycle sequencing with dye terminators. Sequencing accuracy of greater than 99% at 600 bases is routinely achieved on the APB MegaBACE 1000 (see Figure), PE Biosystems ABI 3700, and slab gel DNA sequencers. |
The protocol was developed and optimized based on standard methods. This rapid and automation-friendly protocol clears the lysate by filtration and relies primarily on vacuum for sample processing, delivering 3-4 g of plasmid per well from two deep 96-well blocks within 60 min. The processing time required for more than two plates depends on the number of available vacuum manifolds.
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