Boreal’s electrophoretic extraction technology offers the simplest solution for recovery of high purity DNA up to 1 Mb from cell cultures, or DNA embedded in agarose and PGFE plugs.
- Extract high yields of DNA from 100 kb to over 1 Mb
- Recover highly concentrated and purified DNA from agarose and PFGE plugs
- Eliminate inefficient and time consuming dialysis, electroelution, or agarase digestion protocols
- Recover high purity DNA ready for library construction, optical mapping, and other genetic analysis
High Purity Nucleic Acids From Inhibited Samples
Boreal’s DNA purification technology has been proven to extract the highest quality nucleic acids from challenging samples including the Athabasca oil sands, Atacama desert and Antarctic tundra. Now, access that same standard of yield and purity with high molecular weights between 100 kb and 1 Mb from cell cultures, agarose and PFGE plugs; or environmental, clinical, plant and other tissue samples. No other protocol offers the ease-of-use, high purity and yield, or molecular weight recovery advantages of Boreal’s technology.
Highest Molecular Weight Recovery – In Solution
Boreal’s electrophoretic purification technology avoids manual sample handling and mechanical shearing forces associated with other purification methods enabling extraction of intact DNA fragments up to 1.1 Mb. This unique two-dimensional separation technology concentrates nucleic acids while removing contaminant molecules and enzymatic inhibitors. The purified DNA is recovered in a buffer containing polyethylene glycol which allows for transfer of the highly concentrated product and subsequent downstream analysis.
Specifications
| Parameter | Specification |
|---|---|
| Input | Up to 4 agarose gel plugs containing crude DNA or lysed cell culture |
| Processing Time | Total time: ~60 h (1.5 h hands-on) Lysis: Up to 30 h (sample dependent) – 30 min hands-on Cartridge preparation: 3 h – 1 h hands-on Aurora run time: 30 h (molecular weight dependent) – <5 min hands-on |
| Expected Molecular Weight Recovery | 100 – 1100 kb |
| Capacity | 20 μg |
| Output Volume | 80 uL buffer |
| Buffer | 2x TBE with Polyethylene glycol (PEG) |
Protocol Overview
- Cast cells in agarose slices
- Gently lyse cells within the agarose slices
- Cast 4 slices in agarose in the Aurora cartridge
- Press Go – No additional handling steps required
- After purification, add 30 ul of 50 mM MgCl2 and wait 5-10 minutes
- Pipette out the purified DNA from central well of the cartridge using a wide bore pipette tip. The DNA is ready for use in downstream applications.
References
- Ultra High Molecular Weight DNA Recovery in Buffer Protocol for Aurora Instruments
- Liles et al., Cold Spring Harbor Protocols, 2009
