Author: Hans Günter Gassen

Publisher:

Published: 1982

Total Pages: 268

ISBN-13:

Preface: In the past the chemical and enzymatic synthesis of oligonucleotides of defined sequence had to be left to a few experts. Now, however, with the triester approach, the phosphite method and the solid-support techniques gene fragment synthesis has turned into an easy procedure even for a non-chemist. Due to the elegant chemistry involved, all methods work without sophisticated equipment and are prone to mechanisation and eventual automation. It is hoped that combined chemical-encymatic gene synthesis may become a standard technique in a molecular biology laboratory, such as DNA sequencing or in-vitro recombination of nucleic acides. We omitted chemical RNA synthesis, since this field is developing so rapidly at the moment that one has to refer to the original publications. However, we included enzymatic synthesis of RNA fragments, procedures which already have obtained a high degree of standardisation. Most of the contributions are revised versions of the protocols supplied for the EMBO sponsored course on "Automated Chemical and Enzymic Gene Synthesis", held in Darmstadt, March 21 to April 3, 1982. The protocols were improved on the basis of the experience of 30 student scientists with chemical, biological or medical backgrounds. Previously omitted procedures, such as the wandering spot method for oligonucleotide analysis, were included. In editing the manuscript we encountered problems with the nomenclature of nucleic acid components. In unambiquous cases we favoured a simple description, hoping for example, that oligodeoxynucleotide is always understood to mean oligo-2'-deoxyribonucleotide. This book aims to provide those interested in DNA/RNA research with state-of-the-art methods in the synthesis, purification, and analysis of DNA and RNA fragments. The editors wish to thank the authors for their efforts in preparing manuscripts from the the revised laboratory protocols. We gratefully acknowledge the skill and the patience of Mrs. E. Ronnfeldt in typing the manuscripts. We express our thanks to Verlag Chemie for the friendly and very efficient cooperation.--H.G. Gassen A. Lang--Darmstadt, in July 1982.