Points for attention about glue recovery and cutting

1. The buffer and gel of electrophoresis are newly made. Clean the gel cutting table and clean the blade.
2. Glue cutting is to recycle all the glue at the location of the whole purpose fragment. In order to reduce the volume of glue, relatively thin glue can be used, as long as enough sample can be used, but also can use a thin and wide comb to run glue.
3 about protection, in the plexiglass is enough, wear a protective mask to protect your eyes, if you wear resin glasses. If you are very afraid of ULTRAVIOLET irradiation, or have special requirements for the glue, such as the requirement that the glue does not contain EB, point marker and a ruler with scale can be used to take photos to determine the position of the target strip on the glue and cut the glue.
4. Point marker gel according to the normal procedure, then cut off the marker glue, stain with EB, and take photos with the graduated ruler under uv lamp. Since the relative position between the band to be recycled and marker is known, the position of the target band on the undyed glue can be measured according to the ruler. If it is difficult to judge, a small number of samples can be directly placed on the marker edge points, so that the position can be easily determined.
Other factors affecting the results of electrophoresis experiments:
Agarose: Agarose of different factories and batches has different impurities, which affect THE migration of DNA and the intensity of fluorescent background, and should be used selectively.
Preparation of gel: the buffer added to the gel should be in phase with the electrophoresis tank, and the melted gel should be poured into the plate in time to avoid solidification and agglomeration before pouring. The gel poured into the plate should be free of bubbles that could affect the electrophoretic results.
Electrophoresis buffer: To maintain the ionic strength and pH required for electrophoresis, the electrophoresis buffer should be updated frequently.