Noboru Sasagawa

By Noboru Sasagawa

Plasmid purification is a rather classical experiment, but the technique is still developing for time- and cost- saving. The critical principle is based on the alkaline lysis method, although the following steps have several variations. The needed purities and/or quantities of DNA depend on researches using isolated plasmids, meaning that more reasonable method can be selected in each experiment. For example, a non-alkaline-lysis method such as boiling method is still available. One of the important steps for purifying plasmid is a removal of RNA. Ribonuclease is usually used for removing RNA from plasmid sample. On the other hand, a kind of salts such as lithium and calcium functions to make RNA as a selective precipitate from DNA-RNA mixture. Based on these backgrounds, the technique to purify plasmid DNA has been discussed.

Part of the book: Plasmid

By Noboru Sasagawa

Agarose gel electrophoresis is one of the most fundamental experiment in biochemistry and/or molecular biology, especially in analyzing deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). Many laboratories do agarose gel electrophoresis almost every day. Besides, sometimes we need to prepare tens of agarose gels at a time for training and/or practices of students. In such situations, the more cost-effective way we have, the much more experiments in laboratories/trainings of students we can achieve. Actually, experiments of using agarose can be achieved in a more inexpensive way. In this manuscript, conditions of agarose gel electrophoresis experiment (agarose, buffer, and equipment) are considered, and achievements of such efforts are described.

Part of the book: Analytical Chemistry