A modified one-step procedure for rapid RNA isolation from insect

Abstract

A modified guanidinium isothiocyanate method was used to extract total RNA from two forest insect species Clostera anastomosis and Saperda populnea. The integrity of RNA was demonstrated by the methods of gel electrophoresis and cDNA analysis. Typical A₂₆₀/A₂₈₀ absorbance ratio of the total RNA was in range of 1.8 to 2.0. The size of double strand cDNAs obtained by RT-PCR was more than 2 kb, which indicated that intact mRNA was obtained. The fragments of β-actin and chitinase gene from the RNA of C. anastomosis were obtained by RT-PCR, which indicated that the RNA could be used for other molecular operation. By this procedure, RNAs could be extracted and analyzed by electrophoresis from at least 8 samples within 4 hours. These results showed that this method was time-and cost-saving and effective.

Keywords

RNA isolation / β-actin gene / Clostera anastomosis / Saperda populnea