The Nucleogen 96 Plasmid Purification Kit is designed for isolation of high-purity plasmid or cosmid DNA for use in routine molecular biology applications such as fluorescent and radioactive sequencing and cloning. The Nucleogen 96 Plasmid Purification procedure is performed entirely in 96-well format, reducing sample handling to a minimum. Bacterial cultures can be grown and lysed in the 96-well blocks not provided with the kit.

Principle

Nucleogen 96 Plasmid Purification plates contain a unique silica gel membrane that binds up to DNA in the presence of a high concentration of chaotropic salt, and allows elution in a small volume of low-salt buffer. Nucleogen Plasmid membrane technology eliminates time consuming phenol–chloroform extraction and alcohol precipitation, as well as the problems and inconvenience associated with loose resins and slurries. High-purity plasmid DNA eluted from Nucleogen 96 Plasmid Purification plates is immediately ready to use — there is no need to precipitate, concentrate, or desalt.

Nucleogen 96 Clarification plates are asymmetric filter units specially designed to allow fast clearing of bacterial lysates generated under high concentrations of chaotropic salts. Crude bacterial lysates are vacuum filtered through the Nucleogen 96 Clarification plate eliminating the need for centrifugation. Particle-free lysates flow directly into the wells of the Nucleogen 96 Plasmid Purification plate, ready for purification and elution.


Procedure

Plasmid purification using Nucleogen 96 Plasmid Purification Kits follows a simple bind–wash–elute procedure. First, bacterial cultures are lysed and the lysates are cleared by filtration through Nucleogen 96 Clarification modules. The cleared lysates are then applied to the Nucleogen 96 Plasmid Purification plate where plasmid DNA adsorbs to the silicagel membrane. Impurities are washed away and pure DNA is eluted in a small volume of elution buffer or water.


Applications

Nucleogen 96 Plasmid Purification Kits provide reproducible yields of high-purity DNA suitable for use in most applications, including:

Plasmid-mediated gene silencing
Automated fluorescent sequencing, including capillary sequencing
Radioactive sequencing
Restriction digestion
Screening
Ligation and transformation
Labeling
PCR template preparation
All other exzymatic reactions