琼脂糖凝胶电泳法（agarose gel electrophoresis）.docVIP

琼脂糖凝胶电泳法（agarose gel electrophoresis） 5 * TBE 20ml buffer solution with water to 200ml, the preparation of 0.5 * TBE dilution buffer, set aside. 2. glue preparation: weigh 0.4g agarose, a 200ml conical flask, adding 0.5 50ml * TBE dilution buffer, placed in a microwave oven (or electric heating) to remove all agarose melting, shake, this is the 0.8% agarose gel. In the process of heating from time to time to shake the agarose particles attached to the bottle wall into the solution. When heating should be covered with the sealing film, in order to reduce water evaporation. 3. rubber preparation: organic glass adhesive plaster at both ends of the groove respectively (width 1cm) tightly sealed. Glue tank will be sealed in a horizontal support, insert the sample comb, observe the comb teeth edge should be 1mm about the gap and the adhesive groove bottom surface. Adding to the ethidium bromide agarose liquid cooled to 45-60 DEG C in (EB) solution to the final concentration of 0.5 g /ml (or not added EB gel after electrophoresis, but EB solution with 0.5 g/ml staining). Pipettes from inside agarose gel sealing plaster small melt, to agarose solution after the solidification of the remaining agarose carefully pour in plastic tanks, make the glue to form a uniform layer. Pour glue when the temperature is not too low, otherwise the solidification is not uniform, speed is not too fast, otherwise prone to bubbles. After the glue completely solidified out comb, careful not to damage the hair at the bottom of the gel, and then added to 0.5 x TBE slot dilution buffer to the liquid level just before the rubber sheet surface. Because of the edge effect near the sample tank there will be some upheaval, hinder the buffer into the sample slot, so we should assure that the buffer should be filled in sample cell. 4. sample: 10 L enzyme solution with 2 L * 6 loading liquid mixed with micro pipette carefully add in sample cell. If the DNA content is low, according to the increase in the p