Index.The recycle of the products of the PCR

From 2013.igem.org

The recycle of the products of the PCR

1> Transfer the products of the PCR into a new 1.5ml centrifuge tube, then estimate the amount of the liquid, then add 3 times the volume of the combination solution BB, then mix it.

2> Transmit the solution into the centrifuge cylinder, and put it remaining for 2mins, then centrifuge for 30 secs. Divide it into two steps if necessary.

3>Leave the filtrate in the collecting tube, and mount the centrifuge cylinder on the collecting tube again.

4>Add 700ul washing liquid WB into the centrifuge tube, then centrifuge it for 30secs at 12000rmp.

5>Leave out the filtrate in the collecting tube, and mount the centrifuge tube on the collecting tube again. Add 500ul washing liquid into the surface of the centrifuge tube and centrifuge at the speed of 12000rmp, leave out the filtrate.

6>Mount the centrifuge tube on the collecting tube again. Put it in the room temperature or in the 37’c culturing case for several mins, air it adequately to rid the rest alcohol.

7>Mount the centrifuge tube after airing on the centrifuge tube. Add 20-50ul washing liquid EB or deionized water, keep it remaining in the room temperature for 1min, and wash off the DNA.

8>The DNA must be detected by polyacrylamide gel electrophoresis, or by the spectroghotometer, in order to be used in the following experiments,. Preserve them in the -20’c refrigerator if not used.