from: PCR Troubleshooting: The Essential Guide (Author: Michael L. Altshuler). Caister Academic Press, U.K. (2006)
"A broad road full of wonderful prospects is about to open up before you," my boss said hinting at my impending firing, after taking a look at an empty gel. "A clear example of complete professional degeneracy," he greeted me the next time. He is a fine man, I like him very much. His greetings, congratulations and wishes of good fortune outside his laboratory have goaded me into an idea to compile a PCR troubleshooting guide of a novel type accompanied by a brief consideration of ways to obviate rather than overcome the PCR problems. If you fail at PCR, consult this book. Then try to figure out the most probable cause of your failure. I wouldn't let this guide slip into a full-scale manual, so inevitably it has had to be oriented to a reader with some experience in PCR. If you are a PCR greenhorn, keep several manuals close at hand while reading it. Although there is a considerable overlap with other troubleshooting guides, some of the most obvious advice, the kind that you generally remember and is invariably mentioned in other textbooks and guides, has been deliberately omitted from my book (detailed account of the hot start, the trick of touchdown, commonly accepted rules for primer selection, the significance of contamination, etc.). The advice that you will find in these pages is the sort of advice that is not usually found elsewhere and that is often the most useful read more ...