Description

MagExtractor-PCR & Gel Clean up- provides a simple and reliable method for the rapid purification of DNA fragments from a PCR solution, enzyme solution, or agarose gel slices utilizing magnetic silica beads. This kit is based on binding properties of DNA to a silica surface in the presence of chaotropic agents (1). The purified DNA fragments can be used directly for general molecular biology experiments.

Features

  • Extracts DNA fragments from ≤ 100 µl PCR solutions or enzyme solutions within 5 minutes.
  • Extracts DNA fragment from ≤ 0.3 g agarose gel slices (TAE or TBE) within 15 minutes.
  • Agarose slices can 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 effectively recovered. Small fragments (< 40 bp) can be removed.
  • Purified DNA fragments can be applied to sequencing, restriction enzyme treatment, labeling, ligation, transformation, etc.

Applications

  • Purification of DNA fragments from various solutions
  • Purification of DNA fragments from an agarose gel slice

Storage conditionn

Store at room temperature

Components

Binding Solution 88 ml
Washing Solution 132 ml
Magnetic Beads 8.5 ml

Principle

The selectivity of extracted nucleic acids can be changed by optimization of the binding and washing solutions. MagExtractor -Genome- (Code No. NPK-101) extracts genomic DNA from various specimens (e.g. whole blood, cultured cells or animal tissues etc.). MagExtractor -RNA- (Code No. NPK-201) extracts total RNA from various specimens (e.g. cultured cells or animal tissues). MagExtractor -Plasmid- (Code No. NPK-301) extracts plasmids from E. coli cells, MagExtractor -Viral RNA- (Code No. NPK-401) is a kit for extracting viral RNA from serum or plasma specimens. MagExtractor -Plant Genome- (Code No. NPK-501) is a kit for extracting genomic DNA from various plant specimens (e.g., leaf, cultured cells, etc.). MagExtractor-PCR & Gel Clean up- (Code No. NPK-601) extracts DNA fragments from a PCR solution, enzyme solution, or agarose gel slices.

Application data

Example 1.Purification of DNA fragments from DNA solutions

DNA fragments were purified from PCR products (120 bp, 1 kb, 2kb) and φx174/Hinf I DNA size markers (0.25 µg/µl) <100 µl> by MagExtractor –PCR & Gel clean up–. The purified DNA fragments were analyzed by agarose and acrylamide gels, respectively. As shown in Fig.1, the fragments approx. > 60 bp could be recovered with high yield. On the other hand, the < 50 bp fragments and primer dimers were removed efficiently (Fig. 2).

Fig. 1. Electrophoretic analysis of the purified PCR products from solutions.

1,3,5: Before purification
2,4,6: After purification


Fig. 2. Electrophoretic analysis of the purified fx174/Hinf I DNA size markers from solutions.

M: φx174/Hinf I Marker
1: MagExtractor
2: Company A
3: Company B
4: Company C

Example 1.Purification of DNA fragments from agarose gel slices

DNA fragments were purified from PCR products (120 bp, 1 kb, 2kb) and λ phage DNA (48.5 kb) from gel slices by MagExtractor -PCR & Gel clean up- and other companies’ purification kits. As shown in Fig. 1 and 2, all fragments could be recovered with high yield using MagExtractor -PCR & Gel clean up-. Other companies’ purification kits showed poor yields in the case of the large DNA fragment (48.5 kb) (Fig. 2).

Fig. 1. Electrophoretic analysis of the purified PCR products from gel slices.

1,3,5: Before purification
2,4,6: After purification


Fig. 2. Electrophoretic analysis of the purified λ phage DNA (48.5 kb) from gel slices.

M: λ/HindIII Marker
1: Before purification
2: MagExtractor
3: Company A
4: Company B

References

  • B. Vogelstein and D. Gillespie, Proc. Natl. Acad. Sci. USA. 76: 615-619 (1979)