RT Journal
A1 Itoh, Masayoshi
A1 Kitsunai, Tokuji
A1 Akiyama, Junichi
A1 Shibata, Kazuhiro
A1 Izawa, Masaki
A1 Kawai, Jun
A1 Tomaru, Yasuhiro
A1 Carninci, Piero
A1 Shibata, Yuko
A1 Ozawa, Yasuhiro
A1 Muramatsu, Masami
A1 Okazaki, Yasushi
A1 Hayashizaki, Yoshihide
T1 Automated Filtration-Based High-Throughput Plasmid Preparation System
JF Genome Research 
JO Genome Research 
YR 1999 
FD May 01 
VO 9 
IS 5 
SP 463 
OP 470 
DO 10.1101/gr.9.5.463 
UL http://genome.cshlp.org/content/9/5/463.abstract 
AB Current methods of plasmid preparation do not allow for large capacity automated processing. We have developed an automated high-throughput system that prepares plasmid DNA for large-scale sequencing. This system is based on our previously reported filtration method. In this method, cell harvesting, alkaline lysis, and plasmid purification occur in a single 96-well microtiter plate from which sequence-ready DNA samples are collected. The plates are designed to allow all reagents to be injected from above the wells and the spent reagents to be aspirated from below. This design has enabled us to build a linear process plasmid preparation system consisting of an automated filter plate stacker and a 21-stage automated plasmid preparator. The 96-well plates used are outfitted with glass-filters that trap Escherichia coli before the plates are stacked in the automated stacker. The plates move from the stacker to each of the 21 stages of the preparator. At specific stages, various reagents or chemicals are injected into the wells from above. Finally, the plates are collected in the second stacker. The optimal throughput of the preparator is 40,000 samples in 17.5 hr. Here, we describe a pilot experiment preparing 15,360 templates in 160 specially designed 96-well glass-filter plates. The prepared plasmids were subjected to restriction digestion, DNA sequencing, and transcriptional sequencing.