Agarose LE (Low Electroendosmosis)
- Vendor: Axygen
- Type: Molecular Biology Reagents
Agarose LE from Axygen Bioscience is a high quality molecular biology grade Agarose suitable for analytical and preparative electrophoresis of nucleic acids. Nucleic acid separation with Agarose LE is between 0.2–15 kbp depending on the concentration of Agarose LE.
Applications
- DNA electrophoresis
- Analysis of PCR products
- Separation of restriction endonuclease digests of DNA
- Electrophoresis of RNA
- Protein Electrophoresis such as radial diffusion
Nucleic acid fragments separated with Agarose LE can be blotted to nylon or nitro-cellulose membranes by all standard blotting techniques. Detection with non-radioactive probes, e.g. digoxigenin (DIG) – labeled nucleic acids, does not interfere with the use of Agarose LE.
LE is tested for preparative electrophoresis and isolation of DNA fragments. However, we recommend also our low melting point Agarose LM that is optimized for these applications.
Specifications
| CLARITY 1.5% (NTU) | ≤ 3% |
| ELECTROENDOSMOSIS (EEO) | 0.05 ‐ 0.13 |
| SULFUR AS SO4 | 0.14% |
| GELLING TEMPERATURE (1.5 %) | 36° C (±1.5°C) |
| MELTING TEMPERATURE (1.5 %) | 88° C (±1.5°C) |
| GEL STRENGTH (1%) | 1200 G/CM2 |
| GEL STRENGTH (1.5 %) | 2500 G/CM2 |
| DNASE | NONE DETECTED |
| RNASE | NONE DETECTED |
Storage and Stability
Store in a cool dry place at 15-25°Celcius.
Protocol: Electrophoresis of DNA and RNA
The most commonly used technique for DNA separation is electrophoresis in horizontal agarose gels submerged in either Tris-acetate or Tris-borate buffer.
RNA molecules are separated in denaturing agarose gels containing formaldehyde. RNA gels are submerged in MOPS buffer.
Step 1: Add Agarose LE to a measured volume of buffer in a beaker or flask that is 3 times the volume of solution prepared. The amount of Agarose LE added depends on the desired concentration.
Step 2: Dissolve the Agarose by melting, simply by heating the slurry in a boiling water bath. If you are using a microwave oven, heat on high power and mix gently.
Step 3: Cool the solution to approx. 50°C before pouring.
Efficient separation of DNA fragments of a wide size range can be achieved by adjusting Agarose LE concentration (see table).
| Concentration of Agarose LE % | DNA separation range (kbp) | Size of linear DNA fragment (bp) |
|---|---|---|
| 0.8 | 1 to 15 | 950 |
| 1 | 5 to 10 | 525 |
| 1.25 | 0.3 to 5 | 450 |
| 1.5 | 0.2 to 4 | 400 |
| 1.75 | 0.2 to 2.5 | 300 |
Results
Agarose LE gels in IX TAE buffer A-0.75%, B-1%, and C-1.25&
Marker: 1kb ladder
Electrophoresis conditions: submarine gel, 2 hours 30 min, 4.5 V/cm in 1XTAE
