ABSTRACT
In this chapter we have tried to assemble a few methods that are needed in a variety of PCR-based procedures. These are basic methods common in molecular biology laboratories, and mastering these techniques is absolutely required for developing a successful PCR project. Extraction and purification of plasmid DNA is an essential technique in modern molecular toxicology studies. The isolation of large quantities of plasmid is required for DNA sequencing, TA cloning, subcloning, and many other procedures. There are several methods that may be used to transfect mammalian cells with plasmid DNA including calcium phosphate coprecipitation, DEAE-Dextran, and electroporation. Agarose gel electrophoresis is a standard method used to separate, identify, and purify DNA fragments including PCR products. Agarose is extracted from seaweed and is a linear polymer of D-galactose and L-galactose. When agarose is melted and then allowed to harden, it forms a matrix which serves as a molecular sieve to separate DNA fragments.
