SuperSep™ DNA
SuperSep™ DNA is non-denaturing polyacrylamide gel for double-stranded DNA. The acrylamide concentration and the gel cross-linking density are optimized for the separation of low-molecular-weight DNA. This product is suitable for the separation of 20 to 200 bp DNA.
Product specifications
| Acrylamide concentration | 15％ |
|---|---|
| Plate size | 10 x 10 x 3 (mm) |
| Gel size | 10 x 10 x 1 (mm) |
| Number of wells | 17 wells |
| Well volume | 25 μL/well |
Recommended running buffers
- 1×TBE Buffer
- 1×Tris-Glycine Buffer (25 mmol/L Tris, 192 mmol/L Glycine)
Example：
5 x TBE (Product Number 318-90041)
10 x Tris-Glycine Buffer (Product Number 201-18601)
Example of use 1: Separation of low-molecular-weight DNA
Using SuperSep™ DNA and Tris-Glycine Buffer, 20 to 200 bp DNA was clearly separated (left). Using TBE Buffer, 30 to 200 bp DNA was clearly separated (right). SuperSep™ DNA can be used to excise PCR products up to 200 bp in length, especially from small RNA cloning gels.
Example of use 2: Comparison with agarose gel
When low-molecular-weight DNA is electrophoresed, bands tend to be smeared on agarose gel, but clear separation is obtained on SuperSep™ DNA.
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Comparison with 1% agarose gel
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Comparison with 2.5% agarose gel
Example of use 3: PCR product electrophoresis
Preprocessing protocol
When the PCR product is directly electrophoresed, bands may be smeared due to contamination with low-molecular-weight proteins. In such circumstances, the following preprocessing is recommended:
- After PCR, add and mix an equal volume of mixture of phenol, chloroform, and isoamyl alcohol (25:24:1).
- Centrifuge the mixture at 4°C and 14,000 rpm (18,800 × g) for 5 minutes, and transfer the top layer to a microtube.
- Add the following to the microtube and mix: 1 μL of Ethachinmate, 10 mol/L Ammonium acetate in one-fourth volume of the aqueous layer collected, and 99.5% ethanol in twice the total volume.
- Centrifuge the mixture at 4°C and 14,000 rpm (18,800 × g) for 15 minutes, and remove the supernatant. Wash the precipitate twice with 70% ethanol.
- After drying at no more than 45°C, dissolve the precipitate in 10 μL of sterile water, and mix the solution with 2 μL of 6 × Loading Buffer Triple Dye.
- Apply 6 μL of the mixture to the gel.
Electrophoresis conditions
Buffer for electrophoresis:：25 mmol/L Tris, 192 mmol/L Glycine
Current : 30 mA x 60min (constant current)
Staining : Ethidium bromide staining × 15 minutes
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