Isolation of RNA and RT‐PCR, cloning, and sequencing of noncoding RNAs from fungi*

Abstract

RNomics, the understanding of functional RNAs and their interactions at a genomic level, is of utmost practical and theoretical importance in modern life sciences. To introduce our students to the techniques and promise of this emerging field, a practical class activity for advanced undergraduate students in biochemistry and molecular biology is described. In these exercises, students first identify noncoding RNA from different fungi by computational methods and analyze their transcription regulation signals and splice signals by bioinformatics tools, then isolate total RNA from these fungi, and finally verify these noncoding RNA gene expressions by reverse transcriptase-polymerase chain reaction, cloning, and sequencing. This activity not only introduces students to the concept of RNomics, noncoding RNA, and RNA splicing, but also introduces students to the practice of basic molecular techniques. The natural combination of the genome projects and bioinformatics with modern molecular biology techniques is considered a major advantage of this laboratory course.