MagExtractor® -PCR & Gel Clean up-
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.
of DNA from DNA Solutions after Enzymatic Reactions or
The recovered DNA can be used in various applications including sequencing, restriction endonuclease digestion and ligation.
1. Recovery of
DNA from Solution
Purification was performed with 100 µl 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 µg/µl), in accordance with the protocol for this kit, for recovery in 100 µl 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.
Feature and Advantages:
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%.
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.
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.
- Add the Binding Solution containing chaotropic reagents to the DNA solution. When using an agarose block, adding the Binding Solution will melt it.
- Add Magnetic Silica Beads and mix for the DNA in the sample to adhere to the silica’s surface.
- Wash beads with Washing Solutions and remove impurities including proteins, salts, dNTPs, and primers.
- Elute and recover DNA from the beads with sterile water or low salt buffer, etc.
Manual use of this kit is simplified with a magnetic beads separation stand. B/F separation may be performed using a compact desktop centrifuge instead of a magnetic beads separation stand.
Purification of Single Strand DNA
This kit is not suitable for purifying single strand DNA.
The recovered DNA solution will contain some EtOH. If it is necessary to evaporate the EtOH, it is advisable to heat the DNA-adsorbed magnetic beads at 55°C for 5 minutes prior to elution.
Beads for Purification
The magnetic beads supplied in this kit can adsorb up to about 5 µg of DNA per 30 µl. Upon use, assume it takes 30 µl of magnetic beads to purify 2.5 µg of DNA, and change the scale if necessary.
|MagExtractor -PCR & Gel Clean up||TYB-NPK-601||200 RXN||€320|