PowerPrep™ Gel Extraction Kits

Technology

The procedure involves solubilizing the gel fragment in a chaotropic salt solution to release the DNA followed by binding to a high capacity silica membrane or resin, washing to remove agarose and buffers and eluting the DNA fragments in either TE or water. In both formats, the gel is dissolved in a sodium perchlorate solution that is compatible with gels run in either TAE or TBE buffers. The resulting DNA is ready to use in a variety of applications. The Rapid spin cartridge columns process DNA up to 10 kb in size in less than 30 minutes. The Matrix format purifies fragments up to 50 kb in 45 minutes and allows easy scale up for purifying DNA from large gel fragments.

(Note: The standard protocol is designed for extracting single or double stranded DNA fragments. A modified protocol should be used for purifying supercoiled plasmids from agarose gels.)

Performance Characteristics

Figure 1 demonstrates the efficient recovery of fragments ranging from 1kb to 12 kb using the Marligen PowerPrep™ Express Gel Extraction Kit. Identical results are obtained with the PowerPrep™ Matrix Gel Extraction Kit. The purified fragments can be completely digested with a variety of restriction enzymes (Fig 2).

Fig. 1 A 1kb DNA ladder was run on a 1% TAE agaose gel. Single bands were extracted with the Marligen PowerPrep™ Express Gel Extraction System. After purification, fragments were re-run, demonstrating excellent yields.

Fig. 2 EcoR1 digested plasmid DNA was extracted from a 1% TAE agarose gel with the Marligen PowerPrep™ Matrix Gel Extraction System. The DNA was then digested with BglII (lane 2), DdeI (lane 3), DraI (lane 4), PvuI (lane 5) and RsaI (lane 6). Lane 1 is the EcoR1 digested control.

Each lot of PowerPrep™ Express Spin Cartridges and Matrix Silica Resin are evaluated for recovery and DNA capacity, and the quality of the purified DNA is verified using restriction endonuclease digestion and ligation. All tests are performed from both TAE and TBE agarose gels.