Guidance on how to use AMRESCO agarose

AMRESCO agarose use guide

Product: Agarose

Item No.: 0710 0815 E776

E434 J234 X174

The correct heating method is essential for the full dissolution and coagulation of agarose. Here, we provide the following points as a reference for the correct preparation of agarose gel.

* AMRESCO agarose takes a short time to heat up and dissolve.

* Agarose is not dissolved in TAE or TBE buffer. When heated, the agarose particles hydrate to form a solution. The hydration is time dependent, and the different agarose hydration points are different. AMRESCO agarose has high purity and ultrafine particles, which is very suitable as an experimental carrier for electrophoresis. Compared with other agarose, AMRESCO agarose has a fast hydration speed, and the user does not need to boil for a long time. Otherwise, the glue is thick, which is not conducive to mixing, and is easily broken after gelation.

Preparation of AMRESCO agarose gel

Preparation of 1% agarose gel with 1×TBE solution

  • The volume of the conical flask used to prepare the glue should be 2-4 times the volume of the glue. Slowly add agarose to the buffer and stir while stirring to prevent the agarose from collecting. Record the total weight of the bottle and solution.
  • The microwave oven is heated for 30 seconds at high heat, and the heating time is adjusted according to the volume of the prepared solution. The heating time is related to the microwave oven, the size of the bottle and the agarose concentration.
  • Shake the agarose solution.
  • Heat again on high heat for 30 s and shake the agarose solution.
  • Return the solution to the microwave again and heat to boiling (about 10-35 s) on high heat. The agarose gel may boil vigorously during contact movement, and be careful to avoid burns. After taking it out of the microwave oven, it was cooled at room temperature for 1-2 min, and gently shaken to overflow the bubbles in the liquid.
  • Put it back in the microwave oven, heat it on high heat, let it boil for about 15 s, observe the dissolved state of the agarose, and if there are still particles, repeat this step until all the crystals are dissolved.
  • After the agarose is completely dissolved into a liquid, the total weight is re-weighed, the amount of the evaporated liquid is calculated, the water is added to the original weight, and the liquid is shaken.
  • It is recommended that the glue be cooled to 50-55 ° C for potting, which is conducive to uniform gel pore size and will not damage the glue meter. Gently shake the agarose solution before filling the gel to release the remaining bubbles in the glue.
  • Glue into the glue tank. The thickness of the glue is 3-5mm. After filling the glue, remove the bubbles between the comb holes or the bottom.
  • Place at room temperature (30-45 min) to allow the gel to fully condense.

If you want to prepare gels of different concentrations and different volumes, please pay attention to the following two points:

  • Shake at least 2 times before boiling the gel solution.
  • After the glue boils, observe the degree of glue dissolution every 10-15s (the interval depends on the concentration and volume of the glue).

Guidelines for small fragment resolution using AMRESCO Agaroses

0815
Agarose 3:1 TM
E776
J234
Remarks
<0.5kb
analysis
~2%
Not recommended
4-5%
TBE Buffer
3.5-5%
TAE or TBE Buffer
Agarose SFR fits, Agarose 3:1 is less transparent
<0.5kb
analysis
～3-5%
3-4%
TBE Buffer
3%
TBE Buffer
3-4%
TAE or TBE Buffer
Agarose SFR and Agarose 3:1 are mechanically stronger than Agarose II at the specified concentrations and are not brittle.
<0.5kb
preparation
2-4%
TAE or TBE Buffer
Not recommended
2-5%
TAE or TBE Buffer
The concentration is selected according to the resolution. Low concentrations increase fragment recovery purity, and TBE buffers are not suitable for some commercial recovery kits.
0.5-2.0kb
analysis
1.5-2.5%
TBE Buffer
1.5-2.5%
TBE Buffer
1.5-2.5%
TBE Buffer
Agarose SFR has a long electrophoresis time and is easy to cause fragment dispersion.
0.5-2.0kb
preparation
1.5-2.5%
TAE or TBE Buffer
Not recommended
1.5-2.5%
TAE or TBE Buffer
Agarose II is suitable, more preferably 1kb or more. TBE buffers are not suitable for some commercial recycling kits.
1.0-3.0kb
analysis
1-1.5%
1-1.5%
1%
Agarose 3:1 is suitable for Agarose II gel strength and is not suitable for imprinting experiments.
Agarose SFR has a long electrophoresis time and is easy to cause small fragments to diffuse.

The above data is for reference only, and the specific selection is based on actual test requirements.

For a reference voltage of 5 V/cm and a low melting point agarose electrophoresis for more than one hour, a lower voltage of 3-4 V/cm or an electrophoresis at 4 ° C is selected.

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