Description
This laboratory manual gives a thorough introduction to basic techniques. It is the result of practical experience, with each protocol having been used extensively in undergraduate courses or tested in the authors laboratory. In addition to detailed protocols and practical notes, each technique includes an overview of its general importance, the time and expense involved in its application and a description of the theoretical mechanisms of each step. This enables users to design their own modifications or to adapt the method to different systems. Surzycki has been holding undergraduate courses and workshops for many years, during which time he has extensively modified and refined the techniques described here. 1 General Aspects of DNA Isolation and Purification.- 2 Preparation of Genomic DNA from Animal Cells.- 3 Preparation of Genomic DNA from Plant Cells.- 4 Preparation of Genomic DNA from Bacteria.- 5 Isolation of Plasmid DNA.- 6 Isolation and Purification of RNA.- 7 Isolation of PolyA+ RNA.- 8 Agarose Gel Electrophoresis of DNA.- 9 Preparation of Probes for Hybridization.- 10 Nucleic Acid Hybridization. A Theoretical Consideration.- 11 DNA Transfer and Hybridization.- 12 Northern Transfer and Hybridization.- 13 DNA Cloning. General Consideration.- 14 DNA Cloning – Experimental Procedures.- 15 DNA Sequencing.- 16 PCR Analysis.
