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Violet BioPak® MicroColumns™ Filled with GlasPac™ Resin
BioPak GlasPac filled MicroColumns contain a proprietary glass resin, which binds dsDNA in the presence of chaotropic salt, and is eluted with water. Applications include purifying PCR products from amplification reactions, cleaning-up DNA from enzymatic reactions, extracting DNA from agarose gel slices, capturing DNA from various Tris buffers, isolating DNA from alkaline lysis procedures, and purifying plasmid DNA from bacterial lysates. These purification applications are typically performed prior to cloning, direct sequencing reactions, DNA amplifications, gel electrophoresis, HPLC, mass spectrometry and other techniques. GlasPac-filled MicroColumns, are not recommended for samples under 50bp. Instead, use our DEAE-filled MicroColumns, which bind low base pair DNA more effectively.
General Principles
The target molecule binds to the GlasPac resin in the presence of sodium iodide (#4 Salt). Sodium chloride (#5 Wash) snd ethanol/ddH20 are then added to wash away unwanted substances. Elution only requires water. There is no need for centrifugation steps normally associated with phenol/chloroform extractions or alcohol precipitaitons. After the elution of the target molecule, subsequent steps are not necessary to prepare the DNA for downstream reactions such as enzyme digest, ligations, transformations, sequences, and amplifications.
Advantages
BioPak GlasPac-filled MicroColumn are a rapid and inexpensive technique for isolating and purifying DNA or RNA from various sources. In combination with the alkaline lysis method for the preparation of bacterial lysates, these MicroColumns constitute an effective method for the isolation of plasmid DNA for sequencing and other purposes.
The high speed of the GlasPac-filled MicroColumn purification process increases the potential to recover biologically active DNA. BioPak MicroColumns are easily adapted for various applications using a 200ul hand-held pipettor. And, because they are mounted from a rack onto a pipettor, MicroColumns never need to handled directly, eliminating enzyme contamination.
Technical Information
Matrix: Silica
Particle Size: Proprietary
Pore Size: N/A
Capacity: 6ug
Binding Efficiency: 95-100%
Elution Efficiency: 85-90% above 100bp (with 2 elution steps), or 70% below 100bp (with 2 elution steps).
Buffers (Included in this QuicKit™)
National Scientific's #4 Salt buffer contains 30ml of saturated sodium chloride. In order to prepare the Final Wash Solution, simply add 102ml of ethanol and 68ml ddH20 to the 30ml of #5 Wash concentrate. The Final Wash solution will wash away the residual salt, enzymes, and agarose from the glass/DNA complex, while maintaining the DNA/glass interaction.
Protocol Guidelines
In summary, preparation for cloning quality templates of dsDNA simply requires the following steps. First, the target molecule is bound to the GlasPac resin in the MicroColumn by pipetting up and down with sodium iodide. Next, washing occurs while sodium chloride and ethanol/ddH20 are pipetted up and down in the MicroColumn. Finally, purified nucleic acid is eluted from the GlasPac resin with water, simply pipetting up and down in the same MicroColumn.
Quick-Step Illustrated Protocol for using Violet GlasPac Resin Filled MicroColumns. (Must have Acrobat Reader 5.0 or higher to view. Click here to download Acrobat Reader 5.1)
Violet GlasPac Resin Filled MicroColumn Written Protocol.
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