Direct Lysis Fosmid96 DNA Purification Kit

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Last modified: 2008-07-15 14:57:15

The Direct Lysis Fosmid96 Kit isolates fosmid DNA by direct lysis of a 200-µl overnight culture, eliminating the need for centrifugation of the culture and resuspension of the cell pellet. The kit is designed to isolate fosmid DNA up to 40 kb for rapid preparation of sequencing templates from induced single-copy fosmids. Typical DNA yields from a 200-µl overnight culture of an induced single-copy fosmid are sufficient for six to eight end-sequencing reactions. This kit can also be used with a high-copy plasmid vector of 1-40 kb and will provide sufficient DNA for up to 50 end-sequencing reactions.

Several manufacturers make microplate DeepWell rotors, which can hold tall microplate filter systems or stacks of microplates can be used that will accommodate the above dimensions. These include:

• Kendro (Sorvall) Legend RT with Highplate rotor
• Beckman
• IEC
• Jouan

A vacuum manifold (for 96-well microplates) with 4-cm depth or more inside should be compatible.

Robotics: Filter plates are automation-friendly because of their standard footprint. Also, the plates can be processed with vacuum filtration of positive pressure with little or no modification of existing instrumentation.

See more data (ASM poster 2008, 2.2M PDF)

Last modified: 2008-06-26 10:55:58

Applications

• Rapid, direct-lysis preparation of fosmid DNA for sequencing.

Benefits

• No cell harvesting or resuspension of the cell pellet.
• The rapid protocol expedites preparation of sequencing templates in a 96-well format.
• Inoculation of culture in a 200-µl volume eliminates the need for 2-ml, deep-well growth plates.
• Low profile filter-receiver plate block H57 x L127 x W85 mm.
• Average 700-950 base read lengths.
• Reduce the amount of media used to under 20 ml per plate compared to 120 ml with a standard 1.25-ml prep.
• Protocol complete in as little as 25 minutes.

Last modified: 2008-06-26 10:53:18

Protocols for: Direct Lysis Fosmid96 DNA Purification Kit

(catalogue number FOS84596)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-06-26 11:02:39