Simple and rapid preparation of plasmid template by a filtration method using microtiter filter plates.
We developed a new simple high-throughput plasmid DNA extraction procedure, based on a modified alkaline lysis method, using only one 96-well microtiter glassfilter plate. In this method, cell harvesting, lysis by alkaline and plasmid purification are performed on only one microtiter glassfilter plate. After washing out RNAs or other contaminants, plasmid DNA is eluted by low-ion strength solution, although precipitated chromosomal DNA is not eluted. The plasmid prepared by this method can be applied to sequencing reactions or restriction enzyme cleavage.
Itoh
M
M
Genome Science Laboratory, Tsukuba Life Science Center, The Institute of Physical and Chemical Research (RIKEN), Koyadai 3-1-1, Tsukuba-city, Ibaraki 305, Japan.
Carninci
P
P
Nagaoka
S
S
Sasaki
N
N
Okazaki
Y
Y
Ohsumi
T
T
Muramatsu
M
M
Hayashizaki
Y
Y
D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary; D004259:DNA-Directed DNA Polymerase; D004587:Electrophoresis, Agar Gel / Q000295:instrumentation / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000737:chemistry / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic; D019914:Taq Polymerase; D013698:Templates, Genetic; D014462:Ultrafiltration / Q000295:instrumentation / Q000379:methods
D018076:DNA, Complementary; D011933:Reagent Kits, Diagnostic; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase