High-speed plasmid isolation using 96-well, size-exclusion filter plates.
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 ofplasmid DNA in a 96-well microplate format. This high-speed protocol employs a modified alkaline lysis methodfor 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 PLASMID plates. Starting with bacterial cell pellets, the entire prycedure 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.
Harris
D
D
Millipore, Danvers, MA 01923, USA. dylan_harris@millipore.com
Engelstein
M
M
Parry
R
R
Smith
J
J
Mabuchi
M
M
Millipore
J Leonard
JL
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005820:Genetic Testing / Q000295:instrumentation / Q000379:methods; D023281:Genomics / Q000295:instrumentation / Q000379:methods*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010957:Plasmids / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D017422:Sequence Analysis, DNA
D004269:DNA, Bacterial