MagExtractor -PCR & Gel Clean up- - Cosmo Bio Co.,Ltd.

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MagExtractor -PCR & Gel Clean up-

Catalog No.: TYB-NPK-601
Size: 200RXN
Price: ¥37400
$499
catalog info: Catalog 2012-p198
Storage: RT
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Purpose: For purification of DNA fragments from various solutions and slices of agarose gel
component: Binding Solution 88 ml
Washing Solution 132 ml
Magnetic Beads 8.5 ml

Other:

 Background 
This kit is for purification of high purity DNA from DNA solutions after enzymatic reactions or from agarose gels after electrophoresing. DNA purified with this kit hardly contains any impurities such as proteins or salts, allowing itsuse in various applications including PCR sequencing, restriction endonuclease digestion and ligation. Since 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. Simple extraction is possible at a 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 can be used in various applications such as removal of primers and dNTPs after PCR reactions, deproteinizing treatments (including BAP control afterreactions), dechlorination of DNA samples prior to electroporation and displacement in buffer after enzymatic reactions. With regard to purification of DNA from agarose gels after electrophoresis, this kit uses a stronger chaotropic agent than the Nalgenerally used to melt agarose, allowing the agarose 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.
 Features 
Extracts DNA fragments from 100 ul PCR solutions or enzyme solutions within 5 minitues.
Extracts DNA fragment from 0.3 g agarose gel slices (TAE or TBE) whithin 15 minitues.
Agarose slicescan be melted at room temperature.
Typical yields from solution or gel slices are approximately 60-70%
DNA fragments of approximately 100 bp to 50 kb can be recovered effectively. Small fragments (< 40 bp) can be removed.
Purified DNA fragments can be applied to sequencing, restriction enzyme treatment, labelling, ligation, transformation, etc.
 Application 
Purification of DNA from DNA solutions after enzymatic reactions or agarose gels. 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 ul of each reaction solution generated from PCR using DNA as a template (120 bp, 1 kb and 2 kb) andX 174/Hinf I Marker (0.25 ug / ul), in accordance with the protocol for this kit, for recovery in 100 ul 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 target DNA fragments had a high level of purity, and had primer dimers eliminated in the PCR products (1 kb). Moreover, the limit range of cut off values was estimated at between 40 and 60 bp.
2. Recovery of DNA from the Agarose Gel Block
40 ul of each PCR solution (120 bp, 1 kb and 2 kb) and the 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 ul of sterile water.
After recovery, DNA was detected by electrophoresis.
It is estimated that the recovery of each PCR product was approximately 70-80% and that of DNA at most 60% based on the results of the experiment.
In addition, results show other commercially available kits produced low yield.
In the electrophoretograms, the difference is observed in the mobility of the 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.



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Reference:

•  B. Vogelstein et al., Proc. Natl. Acad. Sci. USA. 76 615-619 (1979)
•  R. Boom, C. et al., J. Clin. Microbiol. 28 495-503 (1990)
 
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