Electrophoresis tanks

The gel cassette is mounted into the electrophoresis apparatus and electrophoresis tanks are filled with electrode buffer (Soln. H). Remove air bubbles at the interfaces between electrode buffer and... 

Gel unit A is assembled with a ready-made gel, and running buffer diluted 10 times with distilled water is added to the electrophoresis tank. 

The nme required for transfer and the voltage used is dependent on the molecular weight and the nature of the protein to be transferred. During transfer, the electrophoresis tank may be cooled with tap water. 

Remove strip from electrophoresis tank, cutting off the ends which had been submerged in the buffer compartments. Allow strip to partially dry (until excess white spirit had dripped off). 

Remove the comb and locate the gel plates in the electrophoresis tank. Add the running buffer. 

Remove slides from tray and transfer carefully to an electrophoresis tank (see Note 16). 

Slides should be placed in a flat-bed electrophoresis tank lengthways with the frosted end towards the anode. It is essential that the tank is level and all slides face the same direction. 

After the gel has solidified, withdraw the gel comb, taking care not to tear the sample wells. Place the gel-casting tray containing the gel in the electrophoresis tank. Add IX TAE to cover the gel completely. 

Remove the comb and the tape when the gel has completely hardened (20-40 min at room temperature). Place the gel in the electrophoresis tank and add enough TBE buffer to the tank to cover the gel (about 1 mm of depth). The top of the wells should be submerged in TBE buffer. 

Remove the tape or mold box and place the mold in an electrophoresis tank and add electrophoresis buffer until the gel is submerged (a few mm). 

Close electrophoresis tank and run samples towards the + pole. Although 5 V/cm is usually used (or even more for minigels), superior results are obtained at 0.5-1.0 V/cm (overnight). 

Recently, Jones et al. (J2) have described the design and operation of a low-volume electrophoresis tank suitable for running ten slab gels (size 150 X 150 X 2 mm). The advantages of this tank are that its construction is relatively simple and its low volume makes it economical to dispense with the electrophoresis buffer after two or three runs. 

J2. Jones, M. L, Massingham, W. E., and Spragg, P., Electrophoresis tank for running multiple slab gels. Anal. Biochem. 107, 446-454 (1980). 

Turn off the power supply and remove the plates from the electrophoresis tank. Disassemble the plates by gently prying off the top plate with a spatula. Cover both sides of the gel evenly and smoothly with SaranWrap film. Mark the sample lanes with a permanent marker. 

Fig. 6.4. High-voltage electrophoresis tank with one cooling plate, (a) Perspective view, (b) cross sectional view. 1, Lid from Plexiglas 2, filter paper sheet 3, thin glass plate bent as an inverted V 4, platinum electrode S, buffer compartment 6, 7, metal tube with small holes 8, outlet of cooling liquid.

Locating reagents (a) ninhydrin (b) isatin (c) Sakaguchi reagent. Apparatus horizontal electrophoresis tank (any horizontal electrophoresis apparatus (e.g. Shandon model U77—Figure 9.3)) could be used in all cases manufacturers provide instructions for the use of their products. 

Apparatus Horizontal electrophoresis tank (in all cases manufacturers provide instructions for the use of their products). 

Remove the comb, fill the electrophoresis tank and load PCR product into comb wells with a mixture of 16 pi of PCR product and 4 pi of load buffer, for good resolution, do not exceed 50 ng of PCR product. 

Pour the electrophoresis buffer into the electrophoresis tank. 

Once the agarose has solidified remove the tape from the casting tray and carefully seat the casting tray in the electrophoresis tank. Remove the comb(s). 

Following the electrophoresis tank manufacturer s recommendations, pour the gel see Section 2.2.). Prepare the IX TAE electrophoresis buffer and add to the tank. Remove the sealing tape from the ends of the gel tray, and submerge the cooled gel in the buffer. Gently remove the comb(s). 

Platinum wire (similar to that from gel electrophoresis tank). 

Horizontal gel electrophoresis tank and power supply (e.g.. Life Technologies, Scotland). 

Rinse the gel sandwich with sterile water (to remove trace amounts of unpolymerized acrylamide) and carefully remove the comb. Assemble the gel sandwich into an electrophoresis tank and All the chambers with lx TBE. Pre-run the gel for 15 min at room temperature before loading the samples. 

Mix 20 4L of second-round PCR products with 5 pL 5X Ficoll-blue loading buffer, and size separate by electrophoresis through 1.5% agarose gels (3.75 g agarose in 250 mL Tris-acelatc/EDTA electrophoresis buffer (TAE) using TAE as buffer in the electrophoresis tank. Visualize PCR products by incorporation of... 

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