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Synergel Instructions and Data Sheet
Step-by-Step Instructions:
Materials Required:
Synergel
Agarose
Ethanol
Buffer: 0.5X TPE Buffer (0.5X) solution: 0.04 Tris base, 0.004M EDTA, pH adjusted to 8.0 with phosphoric acid. Alternatively, 1X TAE or 0.5X TBE may be used.
Instructions:
1. Weigh out the required amount of agarose and place in a dry beaker or flask. Agarose concentration is generally fixed at 0.7% (w/v).
2. Calculate and weigh out the appropriate amount of Synergel, which is generally between the range of 0.2-2.0% (w/v). Higher concentrations of Synergel (3-4%) may be used for fractionating very small molecules (100bp or less). The Synergel concentration is derived by taking the normally used agarose concentration, subtracting 0.7% and multiplying by 0.5.
3. Add the Synergel powder to the beaker or flask and thoroughly intermix with the agarose.
4. Thoroughly saturate the mixed powders with ethanol. (Enough ethanol so that the resulting slurry can move about freely at the bottom of the container.)
5. Gradually add 0.5X TPE buffer or alternative buffer while swirling the container to prevent clumping.
6. Dissolve the suspension in a microwave or by direct heating. Intermittent swirling during the heating process helps to ensure the best dispersal, avoids boil over and facilitates rapid dissolving of the suspension. To be sure that all particles are dissolved, hold the suspension to the light, if no refractile particles are visible, the solution may be cooled and poured. If particles persist, re-microwave to boiling then let solution sit for approximately 30 seconds. Repeating this procedure 2-3 times may be required before particles dissolve into solution. Gelling temperature will be approximately the same as set for the agarose component alone. After the gel has “set” remove the comb and add buffer in preparation for electrophoresis.
Troubleshooting Note:
The use of very clean combs facilitates easy removal of the combs and prevents tearing of the gel walls.
Reference:
Synergel represents a second generation modified polysaccharide product related to the gel composition described in Analytical Biochemistry 163:247-254 (1987).