Restriction Endonucleases

Restriction enzymes are extremely reliable and easy to use, perhaps to the point of their performance being taken for granted. Have you recently evaluated your criteria for selecting an enzyme for your restriction digest? Could there be another enzyme that recognizes your restriction site whose enzymatic properties would benefit your research? Exactly which properties might be the subject of the previous question? What does cost tell you about the quality of an enzyme preparation? The chapter begins with a discussion about the practical aspects of comparing and selecting restriction enzymes most appropriate for your research needs. Once you've selected your enzyme(s), are you prepared to use them in the most efficient manner? Can they be diluted? Is there any practical use to the quality control information supplied with the enzyme? Is it necessary to adhere to the recommended reaction conditions? Does template structure affect enzyme performance? Does it matter if your reaction could be categorized as "simple" or "complex"? What difference would this mean to your digestion conditions? The final section of the chapter discusses the digestion of genomic DNA, including a comparison of strategies and techniques that insert rare cutting sites into genomic DNA.