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Polyacrylamide gel for low-molecular-weight DNA

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.

  • Comparison with 1% agarose gel
  • 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:

  1. After PCR, add and mix an equal volume of mixture of phenol, chloroform, and isoamyl alcohol (25:24:1).
  2. Centrifuge the mixture at 4°C and 14,000 rpm (18,800 × g) for 5 minutes, and transfer the top layer to a microtube.
  3. 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.
  4. 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.
  5. 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.
  6. 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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Loading Buffers

Running Buffers

DNA Markers

Product content may differ from the actual image due to minor specification changes etc.

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