DirectPrep 96 Kits

The DirectPrep 96 Kits are designed for easy, cost-effective, high-throughput purification of high-copy-plasmid DNA. The system uses a simple one-plate procedure and a modified alkaline lysis chemistry that makes clearing of the lysate unnecessary. Up to 96 minipreps are prepared in 60–65 minutes on the BioRobot 3000 and BioRobot 8000 (both no longer available), or in 35 minutes using the QIAvac Multiwell. The purified DNA is suitable for use in standard downstream applications such as sequencing and automated fluorescent sequencing (see figure "High-quality DNA").

The DirectPrep 96 Kits provide DirectPrep 96 Plates, buffers, growth blocks, and other components required for preparation of 4 x 96 plasmid minipreps. The plates are standard format according to the Society for Biomolecular Screening.

The DirectPrep 96 method of high-throughput plasmid DNA isolation uses a novel lysis chemistry that eliminates the need for a lysate clearing step. Plasmid DNA purification is performed using only one plate and a simple bind-wash-elute procedure. This results in a protocol that is easier to perform than most existing methods. The DirectPrep 96 procedure is designed for use on the BioRobot 3000 or BioRobot 8000 workstations and can also be performed manually using the QIAvac Multiwell.

Specifications

Features	DirectPrep 96 BioRobot Kit	DirectPrep 96 MiniPrep Kit
Applications	PCR, fluorescent sequencing, restriction digestion, etc.	PCR, fluorescent sequencing, restriction digestion, etc.
Culture volume/starting material	1.25 ml culture volume	1.25 ml culture volume
Plasmid type	High-copy	High-copy
Processing	Automated	Manual (vacuum)
Samples per run; throughput	96 samples per run	96 samples per run
Technology	Silica technology	Silica technology
Time per run	<65 minutes	35 min/plate
Yield	<4 µg	<4 µg

All lysis steps are carried out in the same 96-well flat-bottom block used for growth of the bacteria. After alkaline lysis of the cells, the lysate is neutralized using an optimized buffer, which leads to a virtually clear lysate. Centrifugation or filtration of the lysate is not required. Isopropanol is added in order to optimize the DNA binding conditions, and lysates are transferred to the activated DirectPrep 96 plate where all subsequent steps take place (see flowchart "DirectPrep 96 procedure"). Plasmid DNA binds to the membrane, which is subsequently washed. After washing, the membrane is dried and the bound plasmid DNA is eluted. The DirectPrep 96 procedure gives typical yields of up to 4 µg DNA from 1.25 ml
E. coli culture grown in LB medium.

The DirectPrep 96 Kits provide ready-to-use plasmid DNA of a yield and quality especially suited for high-throughput applications, including sequencing, PCR, restriction digestion, and transformation.