Silica Bead DNA Gel Extraction Kit

Detailní popis

Thermo Scientific Silica Bead DNA Gel Extraction Kit is a simple and efficient system for DNA extraction from agarose gels and reaction mixtures. The kit utilizes the modified protocol of Vogelstein and Gillespie (see Reference 1), employing solubilization of the agarose gel and selective adsorption of nucleic acids on specially prepared silica particles at high salt concentration. Silica particles bound with DNA are washed to remove contaminants and pure DNA is eluted with TE buffer or water.

This method requires few manipulations, and is both faster and easier to perform than other organic-based extraction methods. Compared to column based gel-extraction kits, the DNA Gel Extraction Kit offers flexibility for scaling reaction and elution volumes up or down. The purified DNA is suitable for all common molecular biology procedures, including restriction digestion, cloning, sequencing, etc.

Highlights

• Flexible – easy to scale up or down both reaction and elution volumes
• Efficient – greater than 80% DNA recovery
• Fast – purification takes only 15 to 20 minutes
• Effective – suitable for DNA fragments as short as 100 bp
• Safe – does not involve phenol extraction

Applications

• Purification of DNA fragments from agarose gels prepared with TAE or TBE buffers
• Concentration of DNA samples for desalting or buffer change
• Removal of enzymes after PCR, restriction digestion, dephosphorylation or labeling reactions
• Removal of unincorporated nucleotides, primers and primer-dimers
• Removal of linkers following ligation
• Removal of residual salts, phenol, chloroform, or ethidium bromide
• Purification of RNA-free plasmid DNA

Includes

• Silica Powder Suspension
• Binding Buffer
• Concentrated Washing Buffer
• TBE Conversion Buffer
• Detailed Protocol

Note

Care should be taken to avoid DNA damage with UV light when the gel-purified DNA fragment will be used in downstream cloning reactions. Always use a long wavelength UV (360 nm) light-box during excision of the gel slice. If only a short-wavelength UV (254 to 312 nm) light-box is available, minimize the UV exposure to a few seconds or keep the gel on a glass or plastic plate. Alternatively, visible dyes can be included in standard agarose gels to allow for visualization of the DNA bands in ambient light (2, 3).