DirectPrep 96 Kits

For high-throughput, automated purification of sequencing-grade DNA

Features

  • Cost-effective purification for high-throughput projects
  • Simple one-plate procedure
  • Automated and manual procedures available
  • Up to 96 samples in just 35 minutes
  • SBS plate standard for easy automation
DirectPrep 96 MiniPrep Kit (4)

Cat. No. / ID: 27361

For 4 x 96 plasmid minipreps: 4 DirectPrep 96 Plates, Buffers, Flat-Bottom Blocks and Lids, AirPore Tape Sheets, Tape Pads, Elution Microtubes RS, Caps
¥81,500
 
DirectPrep 96 Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

DirectPrep 96 Kits provide 96-well purification plates suitable for vacuum or automated processing, yielding sequencing-grade plasmid DNA. These cost-effective Kits are highly suited for sequencing and screening projects. The lysis chemistry eliminates the need for a lysate clearing step.
The product must be used in conjunction with QIAvac Multiwell.

Performance

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.

Principle

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

Procedure

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.

Applications

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.

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPCR, fluorescent sequencing, Restriction digestion etc.
Culture volume/starting material1.25 ml culture volume
ProcessingManual (vacuum)
Samples per run (throughput)96 samples per run
Plasmid typeHigh-copy
TechnologySilica technology
Time per run or prep per run35 min/plate
Yield<4 µg

Resources

Quick-Start Protocols (1)
Kit Handbooks (1)

Publications

Microeukaryote community patterns along an O2/H2S gradient in a supersulfidic anoxic fjord (Framvaren, Norway).
Behnke A; Bunge J; Barger K; Breiner HW; Alla V; Stoeck T;
Appl Environ Microbiol; 2006; 72 (5):3626-36 2006 May PMID:16672511

FAQ

3312 - For DirectPrep 96 miniprep, is it possible to use a half plate and save the other half for later prep?

Yes, it is possible. The unused wells of the plate should be sealed with a tape. However, it is recommended to use more than 2 rows (16 wells) in order to avoid unequal vacuum pressure which leads to low yields.

What is the average DNA concentration obtained using the DirectPrep 96 Miniprep Kit?

The average DNA concentration obtained with the DirectPrep 96 Miniprep Kit is 66 ng/ul (typically >50 ng/ul) in an eluate volume of 50 ul. Up to 4 ug DNA total can be purified from 1.3 ml of LB culture medium. Please note that the actual yield will depend on the vector and host strain being used.

The DirectPrep 96 procedure is suitable for use with high-copy plasmids only.

FAQ ID -721
What is the RNase A concentration and composition of Buffer P1?

The composition of Buffer P1 is:

  • 50 mM Tris·Cl, pH 8.0
  • 10 mM EDTA
  • 100 µg/ml RNase A

After RNase A addition, the buffer should be stored at 2–8°C.

Buffer P1 is the resuspension buffer used in a variety of QIAGEN kits for plasmid DNA purification. Details on buffer preparation and storage are presented in Appendix B of the QIAGEN Plasmid Purification Handbook.

FAQ ID -198
What are Elution Microtube adapters and Microplate adapters used in the DirectPrep 96 Miniprep Kit, and how can I order them?

The Elution Microtube adapter of the DirectPrep 96 Miniprep Kit is an adapter needed to securely hold a rack of Elution Microtubes RS in the QIAvac Multiwell vacuum manifold. You can order the Elution Microtube adapter using part number 9236778.

The Microplate adapter is required to secure 96-well microplates in the QIAvac Multiwell vacuum manifold. The Microplate adapter can be ordered using part number 9236850.

 

 

FAQ ID -646
Can the QIAvac 96 vacuum manifold be used with DirectPrep 96 plates?

No. The DirectPrep 96 plate of the DirectPrep 96 Miniprep Kit cannot be securely positioned in the QIAvac 96 vacuum manifold, and the nozzles may not fit into the correct wells below. Furthermore, adapters are needed to position Elution Microtubes or Microplates in the correct height for elution. If the distance of the "receiving" plate to the nozzles is not optimal, splashing and thus cross contamination can occur. Due to these difficulties with fitting, tightness and potential cross contamination, we do not recommend using the QIAvac 96 with the DirectPrep 96 System.

We recommend to use the QIAvac Mulitwell vacuum manifold in combination with the DirectPrep 96 Miniprep Kit.

FAQ ID -722