This kit is for use in purification of high purity DNA from DNA solutions after enzymatic reactions or agarose gels after electrophoresing. DNA purified with this kit contains hardly any impurities such as proteins or salts, enabling use in various applications including PCR sequencing, restriction endonuclease digestion and ligation. As this kit employs the principle that magnetic silica beads bind genomic DNA present in a lysate solution, it is not necessary to perform deproteinization using harmful reagents such as phenol, ethanol precipitation, or high-speed centrifugation, enabling simple extraction at low cost. Unlike commercially available spin columns using silica gel or matrix, this kit allows free adjustment of process scales according to sample amounts, providing convenient, economic extraction. This kit enables use in various applications such as removal of primers and dNTPs after PCR reactions, deproteinizing treatments including BAP control after reactions, dechlorination of DNA samples prior to electroporation, and displacement in buffer after enzymatic reactions. With regard to the purification of DNA from agarose gels after electrophoresing, this kit uses a stronger chaotropic agent than the Nal generally used to melt agarose, allowing it to melt at room temperature in a short time. This kit is suitable for the MFX Series automatic nucleic acids purification system and can also be used as a manual kit for B/F (solid-liquid) separation using a magnetic beads separation stand.
1. Recovery of DNA from Solution
Purification was performed with 100 l of each reaction solution generated from PCR using λ DNA as a template (120 bp, 1 kb and 2 kb) and ΦX174/Hinf I Marker (0.25 g/l), in accordance with the protocol for this kit, for recovery in 100 l of sterile water.
After recovery, the PCR products and the marker were analyzed by agarose gel electrophoresis and acrylamide electrophoresis, respectively.
The results showed that use of this kit recovered the target DNA fragment at a high level of purity, and had eliminated primer dimer, which was observed in the PCR products (1 kb). Moreover, the limit range of cut off values was estimated at between 40 and 60 bp for this kit as well as other commercially available kits.
2. Recovery of DNA from the Agarose Gel Block
40 µl of each PCR solution (120 bp, 1 kb and 2 kb) and the λ DNA solution (48.5 kb) were electrophoresed using TBE and TAE agarose gels, respectively, and then the target bands were digested and purified for recovery in 40 µl of sterile water.
After recovery, the DNA was detected by electrophoresis.
It is estimated that the recovery of each PCR product was approximately 70-80% and that of λ DNA at most 60% based on the results of the experiment.
In addition, the results show extremely low yield was produced using the other commercially available kits.
In the electrophoretograms, the difference is observed in the mobility of the λ DNA before and after recovery, attributed to the variation in salt concentration of the DNA solution.
The results of other experiments also demonstrated that the recovered DNA can be used in various applications including sequencing, restriction enzyme digestion and ligation.
Efficient Purification from a Variety of Samples
Enables purification of high purity DNA from enzyme reaction solutions or electrophoresed agarose gel with an average recovery rate of more than 70%.
Quick, Simple Extraction
MagExtractor -PCR & Gel Clean up- is based on the principle of DNA adsorbance by magnetic silica beads, and can purify 60bp - 50kb of DNA quickly and simply (DNA solution: approx. 5 min., agarose gel: approx. 15 min.). Purification from agarose gel does not require the use of a heat block or hot water bath as has been required with conventional methods to melt the agarose.
No Phenol or Chloroform Extraction
This kit does not require the use of harmful phenol or chloroform so there is no hazardous waste problem.
This kit provides high purity DNA according to the following steps.
MagExtractor -PCR & Gel Clean up-
To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.