一种原位灌注

【资料简介】

1. Rinse off the minigel apparatus, the gel casting tray and the desired combs with water.

2. Weigh out 0.8 g of agarose for every 1% concentration of gel you desire. Place this in a 250 ml Ehrlenmeyer flask.

A typical minigel is 1%.

3. To <700 ml of DD-H₂O, add 14 ml of 50x TAE buffer and mix.

4. Add 80 ml of your 1x TAE to the agarose.

5. Melt the agarose in a microwave oven as follows:

45 sec @ HIGH/Mix/45 sec @ HIGH/Mix/1 min @ DEFROST/Mix

If the agarose is properly dissolved you should see no globules floating in the solution when held up to the light.

6. Add 4 ul of Ethidium bromide to the solution and slowly pour the gel into the apparatus. Remove any bubbles with a pipet tip if necessary.

7. Let the gel harden, turn the casting tray 90 degrees around, and pour in the remaining buffer solution.

8. Add the proper amount of 6x loading dye to each sample, mix by pipetting up and down, and carefully load each sample into a well using a Pipetman. Also load any molecular weight markers that are needed.

9. Attatch the minigel apparatus cover and the power supply leads, turn on the power supply and adjust the current to the desired level. 100 mAmps is a typical amount of current applied to a minigel, although you may adjust this to speed up the gel or slow it down.