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Abstract for Wizard® Plus Midipreps DNA Purification System
Small-scale purifications of plasmid DNA, known as minipreps, are commonly
used in molecular biology procedures. Over the years many miniprep protocols
have been used, but few have proven to be consistently reliable. The strategy
has been improved and adapted for processing larger culture volumes, 10-100ml,
with the Wizard® Plus Midipreps DNA Purification
System.
The Wizard® Plus Midipreps DNA Purification System
eliminates many of the problems associated with standard midiprep procedures,
and provides a simple and reliable method for rapid isolation of plasmid DNA.
This system can be used to isolate any plasmid, but works most efficiently with
plasmids <20,000bp. When using the standard protocol, the entire midiprep
process can be completed in 90 minutes or less, with no organic extractions or
ethanol precipitations. Multiple midipreps can be easily processed at one time
with the Vac-Man® (20-sample capacity, Cat.# A7231) or Vac-Man®
Jr. (2-sample capacity, Cat.# A7660) Laboratory Vacuum Manifold. DNA is eluted
from the Wizard® Midicolumn in Nuclease-Free Water (Cat.# P1193).
The purified plasmid can be used directly for automated fluorescent DNA
sequencing or restriction digestion without further manipulation, and also can
be used for in vitro transcription reactions supplemented with a ribonuclease
inhibitor, such as Recombinant RNasin®
Ribonuclease Inhibitor (Cat.# N2511).
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