Methods for RNA Isolation, Characterization, and Sequencing
Paul Zumbo and Christopher E. Mason
from: Genome Analysis: Current Procedures and Applications (Edited by: Maria S. Poptsova). Caister Academic Press, U.K. (2014)
Ribonucleic Acid (RNA) is a key substrate for storing and transmitting biological information in cells, along with deoxyribonucleic acid (DNA), proteins, and other small molecules and metabolites. Since the discovery of nucleic acids by Friedrich Miescher in 1869, RNA has been observed in an expanded range of functions within and between cells, tissues, and even between generations. In 1958, Francis Crick proposed the Central Dogma of Molecular Biology, and he placed RNA as a simple intermediary of unidirectional information transfer between DNA and proteins. Yet, today, we know that a wide range of activities surround and violate this dogma, and recent work has shown that RNAs come in many varieties, serve essential regulatory and catalytic roles, and that RNA bases can harbor many small, chemical modifications that can also change its function. This chapter will review the history of RNA's expansion as a mediator and as a catalytic molecule in cells, the new methods developed to characterize and sequence RNA, and the means for contextualizing the roles of RNA read more ...