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DNA Gel Extraction Kit

#K0513	for 100 preps (up to 5 µg of DNA per prep)
Related Documents (in pdf, ~50 KB): Detailed Protocol: #K0513 MSDS (English) MSDS (English-USA) MSDS (German)

 

Features

• Efficient - greater than 80% DNA recover.
• Fast - purification takes only 15-20 min.
• Effective - suitable for DNA fragments as short as 120 bp.
• No toxic reagents used.
• Purified DNA is suitable for direct use in restriction analysis, sequencing, PCR, cloning and labeling.
• Flexible - easy to scale up or down.

DNA extraction from agarose gel.

Description
The DNA Gel Extraction Kit provides a simple and efficient method for DNA extraction from agarose gels and reaction mixtures. The kit utilizes the modified protocol of Vogelstein and Gillespie (1). It is based on the solubilization of 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 the 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. The purified DNA is suitable for all common molecular biology procedures, including restriction digestion, cloning, sequencing, etc.

Applications

• Purification of DNA fragments from any type of agarose gel (TAE or TBE buffers).
• Concentration of DNA samples for desalting or buffer change.
• Removal of proteins after restriction digestion, dephosphorylation, labeling reactions or PCR.
• Removal of unincorporated nucleotides, primers and primer-dimers.
• Removal of linkers after ligation.
• Removal of residual phenol, chloroform or ethidium bromide.
• Purification of RNA-free plasmid DNA.

Quality Control
The kit is tested in purification of 950 bp and 120 bp DNA fragments from 1% agarose gel, and from PCR mixtures as well.

Components of the Kit

• Silica Powder Suspension
• Binding Solution
• Concentrated Washing Buffer
• TBE Conversion Buffer
• Detailed Protocol (in pdf, ~57 KB):
  • #K0513

Note  
For subcloning of gel-purified DNA fragments, care should be taken to avoid DNA damage with UV light. Always use a long wavelength UV (360 nm) light-box during excision of agarose gel slice. If only a short-wavelength UV (254312 nm) light-box is available, minimize the UV exposure to several seconds, or keep the gel either on a glass plate or on a plastic plate, while transilluminating. Alternatively, visible dyes can be included in standard agarose gels to visualize DNA bands in ambient light (2, 3)

Recovery of individual DNA fragments of GeneRuler™ 100 bp Plus DNA Ladder (M) 
from a 1.7% agarose gel in 1X TAE buffer using DNA Gel Extraction Kit

Related Products

TopVision™ LE GQ Agarose TopVision™ LM GQ Agarose 50X TAE Buffer 10X TBE Buffer Calf Intestine Alkaline Phosphatase (CIAP) Shrimp Alkaline Phosphatase (SAP) Rapid DNA Ligation Kit Rapid DNA Ligation & Transformation Kit InsTAclone™ PCR Cloning Kit CloneJET™ PCR Cloning Kit T4 DNA Ligase TransformAid™ Bacterial Transformation Kit FastDigest® Restriction Enzymes PureExtreme® Restriction Enzymes

References

1.  Volgelstein, B., Gillespie, D., Preparative and analytical purification of DNA from agarose, Proc. Nat. Acad. Sci. USA, 615-619, 1979.
2. Rand, K.N., Crystal Violet can be used to Visualize DNA Bands during Gel Electrophoresis and to Improve Cloning Efficiency, Elsevier Trends Journals Technical Tips, Online, T40022, 1996.
3. Adkins, S., Burmeister, M., Visualization of DNA in agarose gels and educational demonstrations, Anal Biochem., 240 (1), 17-23, 1996.

Updated vasario 05, 2008 14:14