DNA Extraction from Agarose Gels
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Overview
Recover 100 to 10,000 bp DNA from agarose gel slices in a single 10-minute spin. The kit consists of a pre-assembled filter device with an agarose gel nebulizer, a microcentrifuge vial, and modified TAE gel extraction buffer (dry).
The device utilizes gel compression to extract DNA from the agarose. Centrifugal force collapses the gel structure, drives the agarose through a small orifice in the gel nebulizer and the resultant gel slurry is sprayed into the sample filter cup.
Prepared DNA requires no further purification for most applications, including cloning and radioisotopic or fluorescent DNA sequencing. Since agarose gel electrophoresis has high resolving power, the small and large non-specific amplification products that frequently interfere with cloning and sequencing after PCR are completely removed from the product.
- Minimal hands-on time
- 10-minute spin
- Fully-functional DNA
Protocol
| Typical DNA Recoveries from Agarose Gels | |
| DNA Size (bp) | Percent DNA Recoveries |
| 100 | 78 |
| 700 | 71 |
| 1000 | 77 |
| 2027 | 47 |
| 4361 | 35 |
| 9416 | 32 |
| 23130 | 29 |
Performance
| 1. | Excise the band of interest after routine electrophoretic analysis of PCR products or other DNA through an agarose gel. |
| 2. | Place the gel slice into the pre-assembled device. Note: Protocol is not compatible with low melting point agarose. |
| 3. | Spin at 5,000 x g for 10 minutes to extract gel slurry into filter cup. |
| 4. | DNA can be stored in the capped filtrate vial after discarding the sample filter cup and gel nebulizer. |
Sequencing Results
Automated fluorescent sequencing results for a pUC18 gel fragment purified with Montage Gel Extraction Device. An ABI PRISM® Dye Terminator Sequencing Ready Reaction Kit with AmpliTaq DNA Polymerase (Applied Biosystems) was used to generate sequencing results. Sequencing results courtesy of Jeff McConnell, Director of Genomic Services, Cleveland Genomics, Cleveland, Ohio.